From a5dfd37509ae28e9a86c7ea3cbc3f548c51aabca Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Tue, 19 Mar 2019 14:44:02 +0100 Subject: [PATCH 01/83] create poppunk --- poppunk_tree | 8 ++++++++ 1 file changed, 8 insertions(+) create mode 100644 poppunk_tree diff --git a/poppunk_tree b/poppunk_tree new file mode 100644 index 0000000..e406e95 --- /dev/null +++ b/poppunk_tree @@ -0,0 +1,8 @@ +# What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) + +[Presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) + + + +# How to use PopPUNK +[PopPUNK Documentation](https://poppunk.readthedocs.io/en/latest/index.html) From 757e0e1b67b860346e60b64fdb19056caa2d5040 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 13:03:51 +0100 Subject: [PATCH 02/83] Create ResFinder.md --- ResFinder.md | 8 ++++++++ 1 file changed, 8 insertions(+) create mode 100644 ResFinder.md diff --git a/ResFinder.md b/ResFinder.md new file mode 100644 index 0000000..ec5e099 --- /dev/null +++ b/ResFinder.md @@ -0,0 +1,8 @@ +**Running ResFinder** + +conda activate ResFinder + +python /usit/abel/u1/jeevka/ResFinder/src/resfinder.py -i -p /usit/abel/u1/jeevka/ResFinder/ResFinderDB/src/ -k /usit/abel/u1/jeevka/ResFinder/KMA/src -o Output + +conda deactivate ResFinder + From b39147b693bdfe70c903bd810eda9d77e82daa82 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 13:04:26 +0100 Subject: [PATCH 03/83] Update ResFinder.md --- ResFinder.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index ec5e099..bc7e099 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -1,8 +1,9 @@ **Running ResFinder** +``` conda activate ResFinder python /usit/abel/u1/jeevka/ResFinder/src/resfinder.py -i -p /usit/abel/u1/jeevka/ResFinder/ResFinderDB/src/ -k /usit/abel/u1/jeevka/ResFinder/KMA/src -o Output conda deactivate ResFinder - +``` From a8183ef8ab4ce0331db1ada2956e21f30245d728 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 13:05:26 +0100 Subject: [PATCH 04/83] Update ResFinder.md --- ResFinder.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/ResFinder.md b/ResFinder.md index bc7e099..df07a5d 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -1,4 +1,6 @@ **Running ResFinder** +---------------------- +use the below code to execute ResFinder Abel. ``` conda activate ResFinder From 964b0ca811b9eb621ee98327380de8a726c8a6c3 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 13:05:49 +0100 Subject: [PATCH 05/83] Update ResFinder.md --- ResFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index df07a5d..f43a2b5 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -1,6 +1,6 @@ **Running ResFinder** ---------------------- -use the below code to execute ResFinder Abel. +Use the below code to execute ResFinder Abel. Contact me if you have any issues. ``` conda activate ResFinder From 3d2e6ccf93c7cdc6b6597b6a5c8828a7c8812b68 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:20:17 +0100 Subject: [PATCH 06/83] Create PlasmidFinder.md --- PlasmidFinder.md | 14 ++++++++++++++ 1 file changed, 14 insertions(+) create mode 100644 PlasmidFinder.md diff --git a/PlasmidFinder.md b/PlasmidFinder.md new file mode 100644 index 0000000..2789c0a --- /dev/null +++ b/PlasmidFinder.md @@ -0,0 +1,14 @@ + +**Running PlasmidFinder** +------------------------- +Use the below code to execute PlasmidFinder Abel. Contact me if you have any issues. + +``` +conda activate PlasmidFinder + +DB="/work/projects/nn9305k/src/PlasmidFinder/PlasmidFinder_DB/plasmidfinder_db/" + +python /work/projects/nn9305k/src/PlasmidFinder/plasmidfinder/plasmidfinder.py -p $DB + +conda deactivate PlasmidFinder +``` From 61965ae4d27d569f7fa880b1f6f1f04529accf70 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:32:38 +0100 Subject: [PATCH 07/83] Update ResFinder.md --- ResFinder.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index f43a2b5..a5d6797 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -1,6 +1,7 @@ **Running ResFinder** ---------------------- -Use the below code to execute ResFinder Abel. Contact me if you have any issues. +Use the below code to execute ResFinder Abel. Dont need to mention BLAST location. +Contact Jeevan if you have any issues. ``` conda activate ResFinder From 2533007c68652410dfb26bb91229970eb8d5436b Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:32:54 +0100 Subject: [PATCH 08/83] Update PlasmidFinder.md --- PlasmidFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 2789c0a..17b0990 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -1,7 +1,7 @@ **Running PlasmidFinder** ------------------------- -Use the below code to execute PlasmidFinder Abel. Contact me if you have any issues. +Use the below code to execute PlasmidFinder Abel. Contact Jeevan if you have any issues. ``` conda activate PlasmidFinder From c510c2fc0cd8f3428bcbf410c9752ca3421c5559 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:48:45 +0100 Subject: [PATCH 09/83] Update PlasmidFinder.md --- PlasmidFinder.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 17b0990..78d8f82 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -1,7 +1,8 @@ **Running PlasmidFinder** ------------------------- -Use the below code to execute PlasmidFinder Abel. Contact Jeevan if you have any issues. +Use the below code to execute PlasmidFinder Abel. Dont need to mention the BLAST location. +Contact Jeevan if you have any issues. ``` conda activate PlasmidFinder From 2d510c4c778442eef3f16446d368f0658a304022 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:53:38 +0100 Subject: [PATCH 10/83] Create VirulanceFiner.md --- VirulanceFiner.md | 14 ++++++++++++++ 1 file changed, 14 insertions(+) create mode 100644 VirulanceFiner.md diff --git a/VirulanceFiner.md b/VirulanceFiner.md new file mode 100644 index 0000000..ec2f21c --- /dev/null +++ b/VirulanceFiner.md @@ -0,0 +1,14 @@ +**Executing VirulanceFinder** +----------------------------- +Don't need to mention about BLAST. Contact Jeevan if there is any issue. + +``` +DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" + +conda activate VirulanceFinder + +python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -d $DB + +conda deactivate VirulanceFinder + +``` From fd88b131de152dae6165605af011c41abc94c011 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:53:58 +0100 Subject: [PATCH 11/83] Update ResFinder.md --- ResFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index a5d6797..85318d1 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -1,4 +1,4 @@ -**Running ResFinder** +**Executing ResFinder** ---------------------- Use the below code to execute ResFinder Abel. Dont need to mention BLAST location. Contact Jeevan if you have any issues. From 28f6f3ec1590f71a1d69b831cebe7182c48304aa Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:54:49 +0100 Subject: [PATCH 12/83] Update PlasmidFinder.md --- PlasmidFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 78d8f82..0409a9b 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -1,5 +1,5 @@ -**Running PlasmidFinder** +**Executing PlasmidFinder** ------------------------- Use the below code to execute PlasmidFinder Abel. Dont need to mention the BLAST location. Contact Jeevan if you have any issues. From be982dc49997abae9f23f6f4c68ca5831dea6362 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:57:33 +0100 Subject: [PATCH 13/83] Create VirulanceFinder.md --- VirulanceFinder.md | 14 ++++++++++++++ 1 file changed, 14 insertions(+) create mode 100644 VirulanceFinder.md diff --git a/VirulanceFinder.md b/VirulanceFinder.md new file mode 100644 index 0000000..ec2f21c --- /dev/null +++ b/VirulanceFinder.md @@ -0,0 +1,14 @@ +**Executing VirulanceFinder** +----------------------------- +Don't need to mention about BLAST. Contact Jeevan if there is any issue. + +``` +DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" + +conda activate VirulanceFinder + +python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -d $DB + +conda deactivate VirulanceFinder + +``` From 0f069c931ca6de3fd6bf188f1270bf1cb5f2a0f1 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 22 Mar 2019 14:57:58 +0100 Subject: [PATCH 14/83] wrong file name no need anymore --- VirulanceFiner.md | 14 -------------- 1 file changed, 14 deletions(-) delete mode 100644 VirulanceFiner.md diff --git a/VirulanceFiner.md b/VirulanceFiner.md deleted file mode 100644 index ec2f21c..0000000 --- a/VirulanceFiner.md +++ /dev/null @@ -1,14 +0,0 @@ -**Executing VirulanceFinder** ------------------------------ -Don't need to mention about BLAST. Contact Jeevan if there is any issue. - -``` -DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" - -conda activate VirulanceFinder - -python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -d $DB - -conda deactivate VirulanceFinder - -``` From ff383ec41b9c36a58cd756f2f94524ba616088e6 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Mon, 25 Mar 2019 13:52:53 +0100 Subject: [PATCH 15/83] Update ResFinder.md --- ResFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index 85318d1..965e322 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -6,7 +6,7 @@ Contact Jeevan if you have any issues. ``` conda activate ResFinder -python /usit/abel/u1/jeevka/ResFinder/src/resfinder.py -i -p /usit/abel/u1/jeevka/ResFinder/ResFinderDB/src/ -k /usit/abel/u1/jeevka/ResFinder/KMA/src -o Output +python /work/projects/nn9305k/src/ResFinder/src/resfinder.py -i -p /work/projects/nn9305k/src/ResFinder/ResFinderDB/src/ -k /work/projects/nn9305k/src/kma/ -o Output conda deactivate ResFinder ``` From eb34b82e6ef976b20ee2ed114e440311fca5ea5e Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Mon, 25 Mar 2019 13:54:09 +0100 Subject: [PATCH 16/83] Update ResFinder.md --- ResFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index 965e322..5431f4e 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -8,5 +8,5 @@ conda activate ResFinder python /work/projects/nn9305k/src/ResFinder/src/resfinder.py -i -p /work/projects/nn9305k/src/ResFinder/ResFinderDB/src/ -k /work/projects/nn9305k/src/kma/ -o Output -conda deactivate ResFinder +conda deactivate ``` From cd0f713e867c5158b361fa4ecdd398fdbc987c78 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Mon, 25 Mar 2019 13:58:53 +0100 Subject: [PATCH 17/83] Update VirulanceFinder.md --- VirulanceFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/VirulanceFinder.md b/VirulanceFinder.md index ec2f21c..5862e27 100644 --- a/VirulanceFinder.md +++ b/VirulanceFinder.md @@ -9,6 +9,6 @@ conda activate VirulanceFinder python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -d $DB -conda deactivate VirulanceFinder +conda deactivate ``` From 66171d79e64440ab0e579ba8ab92413dedcfb7e8 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Mon, 25 Mar 2019 14:28:48 +0100 Subject: [PATCH 18/83] Update VirulanceFinder.md --- VirulanceFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/VirulanceFinder.md b/VirulanceFinder.md index 5862e27..7241f7f 100644 --- a/VirulanceFinder.md +++ b/VirulanceFinder.md @@ -7,7 +7,7 @@ DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" conda activate VirulanceFinder -python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -d $DB +python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -p $DB conda deactivate From b7973c2b0a3cdf9e05500e48f880ed6c020176ae Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Tue, 26 Mar 2019 12:55:14 +0100 Subject: [PATCH 19/83] added PointFinder DB --- ResFinder.md | 3 +++ 1 file changed, 3 insertions(+) diff --git a/ResFinder.md b/ResFinder.md index 5431f4e..4319b1f 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -6,6 +6,9 @@ Contact Jeevan if you have any issues. ``` conda activate ResFinder +# Location of PointFinder DB +PF_DB="/work/projects/nn9305k/src/PointFinder_DB/src/" + python /work/projects/nn9305k/src/ResFinder/src/resfinder.py -i -p /work/projects/nn9305k/src/ResFinder/ResFinderDB/src/ -k /work/projects/nn9305k/src/kma/ -o Output conda deactivate From 1ea57af0ab86bebe1fb8c4e102cf04dcad325fcd Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Tue, 26 Mar 2019 17:39:59 +0100 Subject: [PATCH 20/83] backup --- poppunk_tree | 8 -------- poppunk_tree.md | 26 ++++++++++++++++++++++++++ 2 files changed, 26 insertions(+), 8 deletions(-) delete mode 100644 poppunk_tree create mode 100644 poppunk_tree.md diff --git a/poppunk_tree b/poppunk_tree deleted file mode 100644 index e406e95..0000000 --- a/poppunk_tree +++ /dev/null @@ -1,8 +0,0 @@ -# What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) - -[Presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) - - - -# How to use PopPUNK -[PopPUNK Documentation](https://poppunk.readthedocs.io/en/latest/index.html) diff --git a/poppunk_tree.md b/poppunk_tree.md new file mode 100644 index 0000000..f7e893a --- /dev/null +++ b/poppunk_tree.md @@ -0,0 +1,26 @@ +# What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) : Population Partitioning Using Nucleotide K-mers + +[Manual](https://poppunk.readthedocs.io/) +[PopPUNK Documentation](https://poppunk.readthedocs.io/en/latest/index.html) +[Presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) + +- whole genome (core + assessory) population analysis/clustering +- distinction between isolates : uses k-mer of different length: `mash` to find core and accessory distances between isolates (**pairwise**) +- the distribution of those distances is used to discriminate clusters (defined as strains) of closely related isolates (similarity:both core and accessory) +- clustering of newly added isolates: EXTENDABLE4 - without the need of reanalizing all samples +- maintenance free and auto-reduce database +> aimed for consistent naming clusters between studies, outbreak detection in minutes + + +# How to use PopPUNK + + + + +# What poppunk does and questions: to prepare presentation + + +# Poppunk commands + + +poppunk --fit-model --distances --ref-db --output --full-db --K From dd3ef80768d86493a7acf1ba1e0065c21b0cfcdb Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 28 Mar 2019 13:21:44 +0100 Subject: [PATCH 21/83] Create PointFinder.md --- PointFinder.md | 16 ++++++++++++++++ 1 file changed, 16 insertions(+) create mode 100644 PointFinder.md diff --git a/PointFinder.md b/PointFinder.md new file mode 100644 index 0000000..d914c52 --- /dev/null +++ b/PointFinder.md @@ -0,0 +1,16 @@ +*Executing PointFinder* +========================= +Location of PointFinder. Contact Jeevan if there is any issues. + +``` +/work/projects/nn9305k/src/PointFinder +``` + +``` +conda activate PointFinder + +PointFinder_DB="/work/projects/nn9305k/src/PointFinder_DB/src/" +python PointFinder.py -p $PointFinder_DB + +conda deactivate +``` From 8c92573aaab1b921dab8c24ed31895c12397f4e8 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 28 Mar 2019 13:47:31 +0100 Subject: [PATCH 22/83] Update ResFinder.md --- ResFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index 4319b1f..4ab9d38 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -7,7 +7,7 @@ Contact Jeevan if you have any issues. conda activate ResFinder # Location of PointFinder DB -PF_DB="/work/projects/nn9305k/src/PointFinder_DB/src/" +PF_DB="/work/projects/nn9305k/src/ResFinder/ResFinderDB/src/" python /work/projects/nn9305k/src/ResFinder/src/resfinder.py -i -p /work/projects/nn9305k/src/ResFinder/ResFinderDB/src/ -k /work/projects/nn9305k/src/kma/ -o Output From 2fce735474600b787154922b019cedf290e8f967 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 28 Mar 2019 22:58:57 +0100 Subject: [PATCH 23/83] Create ShigaTyper.md --- ShigaTyper.md | 18 ++++++++++++++++++ 1 file changed, 18 insertions(+) create mode 100644 ShigaTyper.md diff --git a/ShigaTyper.md b/ShigaTyper.md new file mode 100644 index 0000000..a586847 --- /dev/null +++ b/ShigaTyper.md @@ -0,0 +1,18 @@ +ShighaTyper +=========== +Original paper +"In Silico Serotyping Based on Whole-Genome Sequencing Improves the Accuracy of Shigella Identification." + + +Location of Shigatyper +´´´ +/work/projects/nn9305k/src/ShigaTyper/shigatyper +´´´ + +Executing ShigaTyper +==================== +´´´ +conda activate ShigaTyper +python /work/projects/nn9305k/src/ShigaTyper/shigatyper/shigatyper.py +conda deactivate +´´´ From e0b63bde58df941b90ab2f7e5533720eb344b97f Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 28 Mar 2019 23:01:42 +0100 Subject: [PATCH 24/83] Update ShigaTyper.md --- ShigaTyper.md | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/ShigaTyper.md b/ShigaTyper.md index a586847..c732e4c 100644 --- a/ShigaTyper.md +++ b/ShigaTyper.md @@ -5,14 +5,14 @@ Original paper Location of Shigatyper -´´´ +``` /work/projects/nn9305k/src/ShigaTyper/shigatyper -´´´ +``` Executing ShigaTyper ==================== -´´´ +``` conda activate ShigaTyper python /work/projects/nn9305k/src/ShigaTyper/shigatyper/shigatyper.py conda deactivate -´´´ +``` From 2dae08b597a87f1baa93bbd92703edc0616a0bd8 Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Fri, 29 Mar 2019 11:24:29 +0100 Subject: [PATCH 25/83] update poppunk --- figures/refine_poppunk.png | Bin 0 -> 178378 bytes poppunk_tree.md | 201 ++++++++++++++++++++++++++++++++++++- 2 files changed, 197 insertions(+), 4 deletions(-) create mode 100644 figures/refine_poppunk.png diff --git a/figures/refine_poppunk.png b/figures/refine_poppunk.png new file 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zf{7lK9^lijB-w-h;Jn*I0R$$Iv1qa@$Od6i(c<#*a)aFY;rL`L2ooTX`eAS_L$nz4 z3lX}p5gq8>B(sZQQ9llU{Zstx_?I}&b%$XmK__D)X`_lyE_iemtO&>Zl~N?nr+@wR z;L8_vr{$Su9M-X?x51kp)tuz{8wa>O=7xz$R7wg1=uPS!%}tsjqN05$)UEE^DS66$iaH{?i)5WNAUBe) t{^u>i{Va?A@96FShfMnahi|p)dNx_nx6@5Ak-&fFPOHc#$z6N!-vDe>$zuQj literal 0 HcmV?d00001 diff --git a/poppunk_tree.md b/poppunk_tree.md index f7e893a..228fd77 100644 --- a/poppunk_tree.md +++ b/poppunk_tree.md @@ -1,4 +1,5 @@ -# What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) : Population Partitioning Using Nucleotide K-mers +# 1. What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) : Population Partitioning Using Nucleotide K-mers +## 1.1 Summary - Overview [Manual](https://poppunk.readthedocs.io/) [PopPUNK Documentation](https://poppunk.readthedocs.io/en/latest/index.html) @@ -10,17 +11,209 @@ - clustering of newly added isolates: EXTENDABLE4 - without the need of reanalizing all samples - maintenance free and auto-reduce database > aimed for consistent naming clusters between studies, outbreak detection in minutes +- analyse up to 10^4 samples in single step - possible to add new samples +- +# 2. How to use PopPUNK +## 2.1 Create datadase -# How to use PopPUNK +- create a list of your assemblies and other sequences you want to include: you need to have the path of the file included, so you need to create the list from the folder you will run PopPUNK: `ls /*fasta > reference_list.txt` +- +- +poppunk --fit-model --distances --ref-db --output --full-db --K + +## 2.2 Fitting a model: what to look for + + + +

+ +

+ +> trick for seing plot : test! - [ ] use move with bondary with 0 + +> kernet density estimate - > can be used to identify outliers and contamination (program to remove those isolates from DB) > - [ ] where? + + + +- how to do: accessory, core, and combined clusters (see sup material) - [ ] + +### 2.2.1 2D GMM +> equal likelihood contours and decision boundary (within and between cluster-assignments ) + +### 2.2.2 HDBSCAN - +`--dbscan` + +### 2.2.3 Evaluate the chosen model +- silhouette index = measure of how similar an object is to its own cluster (cohesion) compared to other clusters (separation) -> must be close to 1 +- network density must be low (mean good separation between clusters: fewer within than between strains links) + +> **important to look at the network** + +#### 2.2.4 Only if the model is really bad: (otherwise -> go to refine model) + +- possibility to optimize using core distance only (vertical boundary) or accessory only (horizontal boundary) +> when? if core and accessory genomes have independent evolution histories - ex. lots of recombination (blurs in blobs), prophages... +> NB: depending on level problem - can be sufficient to only refine model + +- **modify sketch size** (increase sensibility detection SNPs also increases running time) -> finer differences in %pi (down to single SNPs) --option -[ ] + +## 2.3. refining the model `--refine-model` + +`poppunk --refine-model --distances --ref-db --output --full-db` + For 2D GMM `--K ` OR For HDBSCAN `--dbscan` + +Common optional options: HDBSCAN & 2D GMM +`--pos-shift ` (away from origin) +`--neg-shift ` (towards origin) +`--manual-start ` +`--indiv-refine` allow these boundaries to be placed independently on core/accessory + +

+> need to create a triangular⁵ boundary - move forward and backward FROM starting point (range) +``` +mean0 (x,y) #for within strains blob +mean1 (x,y) #for between strains blob +start (x,y) #starting point to move boundary to +``` +

+ + +example: `poppunk --refine-model --distances lm_example/lm_example.dists --ref-db lm_example_2DGMM --output refine_gmm --full-db --neg-shift 0.1` +to obtain the same refinement as published in their article + + + +NB: **Better to redo all the steps in the same folder - now that we now the best fit - to facilitate further use** + +## 2.4. Simplification database (optional) +When model is good -> then we can stop using `--full-db` option (but not compulsory) + +## 2.5 Visualisation resuts +> output can be made either from --model-fit or --refine-model +`--microreact` Generate output files for microreact visualisation +`--cytoscape `Generate network output files for Cytoscape +`--phandango` Generate phylogeny and TSV for Phandango visualisation +`--grapetree` Generate phylogeny and CSV for grapetree visualisation +`--rapidnj` RAPIDNJ (Path to rapidNJ binary to build NJ tree for Microreact) +`--perplexity `PERPLEXITY (Perplexity used to calculate t-SNE projection (with –microreact) [default=20.0]) +`--info-csv` INFO_CSV (additional metadata: Epidemiological information CSV formatted for microreact (can be used with other outputs)) + + +### 2.5.1 Microreact + +### 2.5.2.GrapeTree + +### 2.5.3. Phandango + +### 2.5.4 Cytoscape + + +## 2.6 Adding new sequences = assigning queries +> addition to the reference network +1. pairwise distances are calculated +2. added as nodes in network to clusters +3. clusters name **DO NOT CHANGE** - unless merged -> then both labels displayed on merged cluster + +`poppunk --assign-query --ref-db --q-files --output --threads <3> --update-db` + +> optional `--model-dir ` if fitted model is in a separated directory +> previsous clustering/network: `--previous-clustering` + +> **OOPS!** for further adding queries: need to use the new database: stored in strain-query `--ref-db ` + +### Queries using core or accessory only; + +--ref-db `--core-only` +--ref-db `--accessory-only ` + + +XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX + -# What poppunk does and questions: to prepare presentation # Poppunk commands -poppunk --fit-model --distances --ref-db --output --full-db --K +# Trics for difficult species: +- Low diversity -> a and %pi not necessary correlated - analyse independently (see supplement) + - genome wide association study: genes responsible differences in a and %pi clusters (pyseer) -> used a-cluster as phenotype + + +# My Points +- [ ] as use random sampling of distances to create network -> maybe important not to unbalanced sampling to create model -> otherwise model risicate to be pushed toward most common lineage (specially if expect new samples different) - same pb as usual ... +- [ ] how to extracti within and between-cluster SNPs distances -> %pi?? +- [ ] understand this sketch size -> per scaffold or over whole genome - must be high!! -> the scetch size -> determines what will be most computationnaly intensive + +- within cluster distance: appear under-estimate - but between comparable to RhierBAPS (maybe depend on model used) + +# ## 2.opening PopPUNK (how does it functions) + +- p_random : maximum probability of random matches kmerse (set to 5% ?? check) -> determines k_min +- kmax set at 29 by default (can increase to max mash is able to support = - [ ] find ) + +## 2.1 Models: classify which distances pairs (%pi and a) are within the same cluster + +### 2.1.1 Model to fit -> Two-dimensional Gaussian mixture model (2D GMM) +Fits random subsample of up to 10⁵ distance pairs +scikit-learn 0.19 +Dirichlet Process prior on weights (nearly 0 to define within-cluster distances) +best final likelihood from five k-means initial starts +K: maximum allowed of mixture components (default = 2) +Distances classified with fitted model + +### 2.1.2 Model to fit -> HDBSCAN (option dbscan) +Classify a subsample of 10⁵ points (distance pairs) using: +Boruvka ball tree Algorithm +Iterations - progressive reductions -> minimum of samples required to initiate the seqrch for a cluster + - -> how conservative clustering is + - -> minimum cluster size (threshold nb points(distances a cluster must contain) + - -> extends points in cluster closest to origin (represent within strains) - should not overlap with between strains clusters + +### 2.2 Networks constructed: +networkx v2.1 -> undirected graph with unweighted egdes -> population clusters +extraction: +- connected components (nodes = samples, connections = pairwise distances) +- ordered by nb isolates (largest to smallest) +Evaluation netwok structure: ns = transitivity (1 - density) +- [ ] transitivity? +- (1-density) -> to subdivise the population + +ns > 0.8 - 1 good fit + +database simplification: randomly select one isolate - per cluster (contected to all other members clusters) + +### 2.3 Refinement distance classification +Models -> treat clusters symmetrically (force clusters same distance/structure) +Refinement -> delimit precisely range %pi - a distances trated as within strain links - to maximize ns + - line between means of within and between strain clusters + - if not correct fit -> provide within and between stran cluster means manually - move boundary starting point (range of move) -> maximize ns (40 equaly spaced points over the allowed range) + +# Summary - Files produced + +| File extension | what | when | +| -------------------------------- | ------------------------------------------------------------------------------------------------------------------------ | --------------------------- | +| *.search.out | pairwise core & accessory distances | | +| *graph.gpickle | network used to predict clusters | | +| *DPGMM_fit.png | scatter plot of all distances, and mixture model fit and assignment | | +| *DPGMM_fit_contours.png | contours of likelihood function fitted to data | | +| *distanceDistribution.png | scatter plot of the distance distribution fitted by the model + kernel-density estimate | | +| *.csv | isolate names and the cluster assigned | | +| *(db).png | unclustered distribution of distances used in the fit (subsampled from total) | | +| *.npz | save fit parameters | | +| *refs | representative references in the new database | (unless --full-db was used) | +| *dbscan.png | scatter plot of all distances, and DBSCAN assignment. | --dbscan | +| *external_clusters.csv | CSV file relating the samples to previous clusters provided in the input CSV. | --external-clustering | +| *core_dists.csv | matrix of pairwise core distances | --microreact | +| *acc_dists.csv | matrix of pairwise accessory distances | --microreact | +| *core_NJ_microreact.nwk | neighbour joining tree using core distances (for microreact) | --microreact | +| *perplexity5.0_accessory_tsne.dot | t-SNE embedding of accessory distances at given perplexity | --microreact | +| *microreact_clusters.csv | cluster assignments plus any epi data added with the --info-csv option (for microreact) | --microreact | +| *cytoscape.csv | cluster assignments plus any epi data added with the --info-csv option (for cytoscape) | --cytoscape | +| *cytoscape.graphml | XML representation of resulting network (for cytoscape) | --cytoscape | +| *refined_fit.png | plot of the new linear boundary, and core and accessory distances coloured by assignment to either side of this boundary | --fit-model | +| *refined_fit.npz | **The saved parameters of the refined fit.** | --fit-model | +| *clusters.csv + .gpickle | for core and accessory |--fit-model --indiv-refine | From 62bc922481523c54d6f9ddd0b680ea6fc1998065 Mon Sep 17 00:00:00 2001 From: Karin Lagesen Date: Mon, 1 Apr 2019 11:32:26 +0200 Subject: [PATCH 26/83] Update README.md Added visualisation exercise. --- README.md | 1 + 1 file changed, 1 insertion(+) diff --git a/README.md b/README.md index 2f15fa6..f73afaf 100644 --- a/README.md +++ b/README.md @@ -20,6 +20,7 @@ The instructors are: * [Use of mash](mash.md) * [Use of conda](conda.md) + * [Mapping and visualization](assembly_visualization.md) ## Course pages From 2f18b5ff9bf969f9bba2a9a17d33a78d8d15a5fd Mon Sep 17 00:00:00 2001 From: Thomas Haverkamp Date: Mon, 1 Apr 2019 12:24:30 +0200 Subject: [PATCH 27/83] Figure is not correctly formated in the document, for now I commented it out --- assembly_visualization.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/assembly_visualization.md b/assembly_visualization.md index fb6667f..45dd5e7 100644 --- a/assembly_visualization.md +++ b/assembly_visualization.md @@ -15,11 +15,12 @@ It is simply finding a/the matching locus/area of a read on a sequence. You coul Usually few mismatches are allowed (think about the consequences). Reads can be mapped as paired or single. If paired is used, then the matching regions are defined by the insert size and the length of each read - +``` +``` ### 1.2 Why mapping reads From fa7b68809d45df9684bf3cefd8ffc8d8819713af Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Mon, 1 Apr 2019 13:14:05 +0200 Subject: [PATCH 28/83] corrected fig format to md obviously does not take a bit more advanced formating back to simple markdown --- assembly_visualization.md | 13 +++---------- 1 file changed, 3 insertions(+), 10 deletions(-) diff --git a/assembly_visualization.md b/assembly_visualization.md index 45dd5e7..05ace9b 100644 --- a/assembly_visualization.md +++ b/assembly_visualization.md @@ -15,12 +15,8 @@ It is simply finding a/the matching locus/area of a read on a sequence. You coul Usually few mismatches are allowed (think about the consequences). Reads can be mapped as paired or single. If paired is used, then the matching regions are defined by the insert size and the length of each read -``` -

- -
-

-``` + +![https://commons.wikimedia.org/wiki/File:Mapping_Reads.png](https://upload.wikimedia.org/wikipedia/commons/2/2e/Mapping_Reads.png) ### 1.2 Why mapping reads @@ -171,10 +167,7 @@ In `Bifrost` we annotated the assembly with `Prokka` (using annotations derived ## 2.4 Loading files in [IGV](https://software.broadinstitute.org/software/igv/) -

- -
-

+![IGV](./figures/IGV.png) 1. Create a `genome file` this allows associating tracks to the assembly : `Genomes > create.genome file`. Use the menu to select your assembly file `.fasta`and the annotation-gene file: `.gff` From 07901cbf304279e980a3593fada36db423997ec2 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 13:46:19 +0200 Subject: [PATCH 29/83] Create SeqSero.md --- SeqSero.md | 12 ++++++++++++ 1 file changed, 12 insertions(+) create mode 100644 SeqSero.md diff --git a/SeqSero.md b/SeqSero.md new file mode 100644 index 0000000..06cd3ed --- /dev/null +++ b/SeqSero.md @@ -0,0 +1,12 @@ +**Excuting SeqSero** +===================== + +*GitHub* +https://github.com/denglab/SeqSero/ + +``` +conda activate SeqSero_Shared +cd /work/projects/nn9305k/src/SeqSero/SeqSero/ +python SeqSero.py --help +conda deactivate +``` From 5cafa700993e1fb4b9c51d697d691d9bbfb2d310 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 14:24:08 +0200 Subject: [PATCH 30/83] Create SeroTypeFinder.md --- SeroTypeFinder.md | 11 +++++++++++ 1 file changed, 11 insertions(+) create mode 100644 SeroTypeFinder.md diff --git a/SeroTypeFinder.md b/SeroTypeFinder.md new file mode 100644 index 0000000..cbb77fb --- /dev/null +++ b/SeroTypeFinder.md @@ -0,0 +1,11 @@ +**Executing SeroTypeFinder** +============================ +https://bitbucket.org/genomicepidemiology/serotypefinder/src/master/ + +``` +conda activate SeroTyperFinder +DB=/work/projects/nn9305k/src/SeroTypeFinder/serotypefinder_db/ +cd /work/projects/nn9305k/src/SeroTypeFinder/ +python serotypefinder.py --help +conda deactivate +``` From d0bfa22d161b356ec3624e119c891ffe7e444670 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:31:21 +0200 Subject: [PATCH 31/83] Update PlasmidFinder.md --- PlasmidFinder.md | 3 +++ 1 file changed, 3 insertions(+) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 0409a9b..f90d559 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -4,6 +4,9 @@ Use the below code to execute PlasmidFinder Abel. Dont need to mention the BLAST location. Contact Jeevan if you have any issues. +**The following code should be used inside a SLURM script** +We are preparing a full help page on how to run using SLURN script. + ``` conda activate PlasmidFinder From 0a3a7a36f10f03ba94ccbfb11621d659a17036e9 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:31:59 +0200 Subject: [PATCH 32/83] Update PointFinder.md --- PointFinder.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/PointFinder.md b/PointFinder.md index d914c52..df34e3f 100644 --- a/PointFinder.md +++ b/PointFinder.md @@ -2,6 +2,11 @@ ========================= Location of PointFinder. Contact Jeevan if there is any issues. + + +**The following code should be used inside a SLURM script +We are preparing a full help page on how to run using SLURN script.** + ``` /work/projects/nn9305k/src/PointFinder ``` From 25714715c60b6d2f00c18ecfaaaf20af76754356 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:32:26 +0200 Subject: [PATCH 33/83] Update ResFinder.md --- ResFinder.md | 4 ++++ 1 file changed, 4 insertions(+) diff --git a/ResFinder.md b/ResFinder.md index 4ab9d38..7725cde 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -3,6 +3,10 @@ Use the below code to execute ResFinder Abel. Dont need to mention BLAST location. Contact Jeevan if you have any issues. + +**The following code should be used inside a SLURM script +We are preparing a full help page on how to run using SLURN script** + ``` conda activate ResFinder From 41495ef8aa40025c7e5b2bcc787ffd80b960569f Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:32:55 +0200 Subject: [PATCH 34/83] Update ResFinder.md --- ResFinder.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/ResFinder.md b/ResFinder.md index 7725cde..df6ff1f 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -4,8 +4,8 @@ Use the below code to execute ResFinder Abel. Dont need to mention BLAST locatio Contact Jeevan if you have any issues. -**The following code should be used inside a SLURM script -We are preparing a full help page on how to run using SLURN script** +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURN script** ``` conda activate ResFinder From 81d59ab37d2406fe29ec9866900fdca23eb59558 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:33:09 +0200 Subject: [PATCH 35/83] Update ResFinder.md --- ResFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/ResFinder.md b/ResFinder.md index df6ff1f..6125044 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -5,7 +5,7 @@ Contact Jeevan if you have any issues. **The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURN script** +**We are preparing a full help page on how to run using SLURM script** ``` conda activate ResFinder From d90cebe50cb906117cc052ebe0b200f0053fe661 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:33:26 +0200 Subject: [PATCH 36/83] Update PlasmidFinder.md --- PlasmidFinder.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index f90d559..d36dd73 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -4,8 +4,8 @@ Use the below code to execute PlasmidFinder Abel. Dont need to mention the BLAST location. Contact Jeevan if you have any issues. -**The following code should be used inside a SLURM script** -We are preparing a full help page on how to run using SLURN script. +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURM script** ``` conda activate PlasmidFinder From edce57f56578faf29864f182d5e25e58403e386c Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:34:05 +0200 Subject: [PATCH 37/83] Update PointFinder.md --- PointFinder.md | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/PointFinder.md b/PointFinder.md index df34e3f..fbd65f4 100644 --- a/PointFinder.md +++ b/PointFinder.md @@ -4,8 +4,9 @@ Location of PointFinder. Contact Jeevan if there is any issues. -**The following code should be used inside a SLURM script -We are preparing a full help page on how to run using SLURN script.** + +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURM script** ``` /work/projects/nn9305k/src/PointFinder From 646c192bc117683f58a7e54c554a4bad2f445a31 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:34:38 +0200 Subject: [PATCH 38/83] Update SeqSero.md --- SeqSero.md | 4 ++++ 1 file changed, 4 insertions(+) diff --git a/SeqSero.md b/SeqSero.md index 06cd3ed..6a1bad8 100644 --- a/SeqSero.md +++ b/SeqSero.md @@ -4,6 +4,10 @@ *GitHub* https://github.com/denglab/SeqSero/ + +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURM script** + ``` conda activate SeqSero_Shared cd /work/projects/nn9305k/src/SeqSero/SeqSero/ From 07205b3c82dbc0a6a32542d20a3275b6d8136d01 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:35:29 +0200 Subject: [PATCH 39/83] Update SeroTypeFinder.md --- SeroTypeFinder.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/SeroTypeFinder.md b/SeroTypeFinder.md index cbb77fb..2936461 100644 --- a/SeroTypeFinder.md +++ b/SeroTypeFinder.md @@ -2,6 +2,11 @@ ============================ https://bitbucket.org/genomicepidemiology/serotypefinder/src/master/ + +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURM script** + + ``` conda activate SeroTyperFinder DB=/work/projects/nn9305k/src/SeroTypeFinder/serotypefinder_db/ From e35438bff678070284527c9373b609d1105ade3b Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:36:09 +0200 Subject: [PATCH 40/83] Update ShigaTyper.md --- ShigaTyper.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/ShigaTyper.md b/ShigaTyper.md index c732e4c..5ae6e7d 100644 --- a/ShigaTyper.md +++ b/ShigaTyper.md @@ -3,6 +3,8 @@ ShighaTyper Original paper "In Silico Serotyping Based on Whole-Genome Sequencing Improves the Accuracy of Shigella Identification." +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURM script** Location of Shigatyper ``` From 14924b06f5725e73a3403768b1719042033e534f Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Thu, 4 Apr 2019 15:36:37 +0200 Subject: [PATCH 41/83] Update VirulanceFinder.md --- VirulanceFinder.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/VirulanceFinder.md b/VirulanceFinder.md index 7241f7f..4a74e57 100644 --- a/VirulanceFinder.md +++ b/VirulanceFinder.md @@ -2,6 +2,11 @@ ----------------------------- Don't need to mention about BLAST. Contact Jeevan if there is any issue. + +**The following code should be used inside a SLURM script.** +**We are preparing a full help page on how to run using SLURM script** + + ``` DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" From e0b18232593ffafcdf3c8b37c7619265d6ddb533 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:21:31 +0200 Subject: [PATCH 42/83] Update PlasmidFinder.md --- PlasmidFinder.md | 30 ++++++++++++++++++++++++------ 1 file changed, 24 insertions(+), 6 deletions(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index d36dd73..cb8d9ee 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -1,18 +1,36 @@ - -**Executing PlasmidFinder** +**PlasmidFinder** ------------------------- +PlasmidFinder identifies plasmids in total or partial sequenced isolates of bacteria Use the below code to execute PlasmidFinder Abel. Dont need to mention the BLAST location. Contact Jeevan if you have any issues. -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** +### For further reading** +https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068535/ +### Here is the SLURM script for Abel to excute the tool. ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=10:00:00 +#SBATCH --mem-per-cpu=32G +#SBATCH --mail-type=ALL + +## Set up job environment: +source /cluster/bin/jobsetup + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + +# Activate Conda environment conda activate PlasmidFinder +# Database location DB="/work/projects/nn9305k/src/PlasmidFinder/PlasmidFinder_DB/plasmidfinder_db/" - python /work/projects/nn9305k/src/PlasmidFinder/plasmidfinder/plasmidfinder.py -p $DB -conda deactivate PlasmidFinder +# deactivate Conda environment +conda deactivate ``` From c98ca3e90b0e08db5cd0485a6247a511880666ef Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:27:27 +0200 Subject: [PATCH 43/83] Update PlasmidFinder.md --- PlasmidFinder.md | 15 ++++++++++----- 1 file changed, 10 insertions(+), 5 deletions(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index cb8d9ee..6f929ad 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -1,13 +1,18 @@ **PlasmidFinder** ------------------------- -PlasmidFinder identifies plasmids in total or partial sequenced isolates of bacteria -Use the below code to execute PlasmidFinder Abel. Dont need to mention the BLAST location. -Contact Jeevan if you have any issues. +PlasmidFinder identifies plasmids in total or partial sequenced isolates of bacteria. -### For further reading** +Dont need to mention the BLAST location. + +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). + +#### User Manual +https://bitbucket.org/genomicepidemiology/plasmidfinder/src + +#### For further reading https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068535/ -### Here is the SLURM script for Abel to excute the tool. +#### Here is the SLURM script for Abel to excute the tool. ``` #!/bin/bash #SBATCH --job-name=DontKillMe From b17c912d16b4193b52563f04622c969bb06ee3d9 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:29:19 +0200 Subject: [PATCH 44/83] Update PlasmidFinder.md --- PlasmidFinder.md | 9 +++------ 1 file changed, 3 insertions(+), 6 deletions(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 6f929ad..1ebfeff 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -12,21 +12,18 @@ https://bitbucket.org/genomicepidemiology/plasmidfinder/src #### For further reading https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068535/ -#### Here is the SLURM script for Abel to excute the tool. +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. ``` #!/bin/bash #SBATCH --job-name=DontKillMe #SBATCH --account=nn9305k -#SBATCH --time=10:00:00 +#SBATCH --time=01:00:00 #SBATCH --mem-per-cpu=32G #SBATCH --mail-type=ALL ## Set up job environment: source /cluster/bin/jobsetup -## Set up job environment: -source /cluster/bin/jobsetup - module load Miniconda3/4.4.10 # Activate Conda environment @@ -34,7 +31,7 @@ conda activate PlasmidFinder # Database location DB="/work/projects/nn9305k/src/PlasmidFinder/PlasmidFinder_DB/plasmidfinder_db/" -python /work/projects/nn9305k/src/PlasmidFinder/plasmidfinder/plasmidfinder.py -p $DB +python /work/projects/nn9305k/src/PlasmidFinder/plasmidfinder/plasmidfinder.py -p $DB -i input_file -o output_file # deactivate Conda environment conda deactivate From d1118037bdcf387e284059b0e595c7c457865a43 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:29:59 +0200 Subject: [PATCH 45/83] Update PlasmidFinder.md --- PlasmidFinder.md | 1 - 1 file changed, 1 deletion(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 1ebfeff..711c104 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -19,7 +19,6 @@ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068535/ #SBATCH --account=nn9305k #SBATCH --time=01:00:00 #SBATCH --mem-per-cpu=32G -#SBATCH --mail-type=ALL ## Set up job environment: source /cluster/bin/jobsetup From 312c3d70a49baeb49261c86ea313d78a9e7fd1c6 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:31:07 +0200 Subject: [PATCH 46/83] Update PlasmidFinder.md --- PlasmidFinder.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 711c104..b3f6c7c 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -2,8 +2,6 @@ ------------------------- PlasmidFinder identifies plasmids in total or partial sequenced isolates of bacteria. -Dont need to mention the BLAST location. - Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). #### User Manual @@ -30,6 +28,8 @@ conda activate PlasmidFinder # Database location DB="/work/projects/nn9305k/src/PlasmidFinder/PlasmidFinder_DB/plasmidfinder_db/" + +# Note: Dont need to mention the BLAST location python /work/projects/nn9305k/src/PlasmidFinder/plasmidfinder/plasmidfinder.py -p $DB -i input_file -o output_file # deactivate Conda environment From 59a302f81b086c1d87f09bd54c0ec11c5d8b4e29 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:39:07 +0200 Subject: [PATCH 47/83] Update PlasmidFinder.md --- PlasmidFinder.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index b3f6c7c..7f66790 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -11,6 +11,11 @@ https://bitbucket.org/genomicepidemiology/plasmidfinder/src https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068535/ #### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory + ``` #!/bin/bash #SBATCH --job-name=DontKillMe From b07cc55969d98c1e1ce50de75af944b90b6f901b Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:47:24 +0200 Subject: [PATCH 48/83] Update PointFinder.md --- PointFinder.md | 34 ++++++++++++++++++++++++---------- 1 file changed, 24 insertions(+), 10 deletions(-) diff --git a/PointFinder.md b/PointFinder.md index fbd65f4..ae39e5a 100644 --- a/PointFinder.md +++ b/PointFinder.md @@ -1,22 +1,36 @@ -*Executing PointFinder* -========================= -Location of PointFinder. Contact Jeevan if there is any issues. +**Executing PointFinder** +------------------------- +The tool detects chromosomal mutations predictive of drug resistance based on WGS data. +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). +#### User Manual +https://bitbucket.org/genomicepidemiology/pointfinder +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** - -``` -/work/projects/nn9305k/src/PointFinder ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=01:00:00 +#SBATCH --mem-per-cpu=32G + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + +# Database location -``` conda activate PointFinder PointFinder_DB="/work/projects/nn9305k/src/PointFinder_DB/src/" -python PointFinder.py -p $PointFinder_DB +python PointFinder.py -p $PointFinder_DB -i input_file -o output_file conda deactivate ``` From aaec97c51af892cb235eb9e134e8dc3e7142ef9b Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:48:00 +0200 Subject: [PATCH 49/83] Update PointFinder.md --- PointFinder.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/PointFinder.md b/PointFinder.md index ae39e5a..18f0bb1 100644 --- a/PointFinder.md +++ b/PointFinder.md @@ -25,10 +25,9 @@ source /cluster/bin/jobsetup module load Miniconda3/4.4.10 -# Database location - conda activate PointFinder +# Database location PointFinder_DB="/work/projects/nn9305k/src/PointFinder_DB/src/" python PointFinder.py -p $PointFinder_DB -i input_file -o output_file From fe85ca811cd33ef17acb4e6d1694456220b340ff Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:50:17 +0200 Subject: [PATCH 50/83] Update ResFinder.md --- ResFinder.md | 24 ++++++++++++++++++++++-- 1 file changed, 22 insertions(+), 2 deletions(-) diff --git a/ResFinder.md b/ResFinder.md index 6125044..a388a20 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -3,11 +3,31 @@ Use the below code to execute ResFinder Abel. Dont need to mention BLAST location. Contact Jeevan if you have any issues. +PlasmidFinder identifies plasmids in total or partial sequenced isolates of bacteria. -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). + +#### User Manual +https://bitbucket.org/genomicepidemiology/pointfinder + +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=01:00:00 +#SBATCH --mem-per-cpu=32G + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + conda activate ResFinder # Location of PointFinder DB From 65e768f5335b0a5bcb468fa53733f453403847db Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 10:54:13 +0200 Subject: [PATCH 51/83] Update SeqSero.md --- SeqSero.md | 32 +++++++++++++++++++++++++------- 1 file changed, 25 insertions(+), 7 deletions(-) diff --git a/SeqSero.md b/SeqSero.md index 6a1bad8..3b73740 100644 --- a/SeqSero.md +++ b/SeqSero.md @@ -1,16 +1,34 @@ **Excuting SeqSero** -===================== +-------------------- +SeqSero is a pipeline for Salmonella serotype determination from raw sequencing reads or genome assemblies. -*GitHub* -https://github.com/denglab/SeqSero/ +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). +#### User Manual +https://github.com/denglab/SeqSero -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** +#### For further reading +http://jcm.asm.org/content/early/2015/03/05/JCM.00323-15 + +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=01:00:00 +#SBATCH --mem-per-cpu=32G + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + conda activate SeqSero_Shared -cd /work/projects/nn9305k/src/SeqSero/SeqSero/ -python SeqSero.py --help +python /work/projects/nn9305k/src/SeqSero/SeqSero/SeqSero.py -m "1" -i input_file -b "mem" conda deactivate ``` From b9a3bed8acaa9d851aa478d73ad2dfd3b60b7ca9 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 11:18:52 +0200 Subject: [PATCH 52/83] Update SeroTypeFinder.md --- SeroTypeFinder.md | 27 ++++++++++++++++++++++----- 1 file changed, 22 insertions(+), 5 deletions(-) diff --git a/SeroTypeFinder.md b/SeroTypeFinder.md index 2936461..18e000a 100644 --- a/SeroTypeFinder.md +++ b/SeroTypeFinder.md @@ -1,16 +1,33 @@ **Executing SeroTypeFinder** ============================ -https://bitbucket.org/genomicepidemiology/serotypefinder/src/master/ +SerotypeFinder identifies the serotype in total or partial sequenced isolates of E. coli. +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** +#### User Manual +https://bitbucket.org/genomicepidemiology/serotypefinder/src/master/ +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory + ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=01:00:00 +#SBATCH --mem-per-cpu=32G + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + conda activate SeroTyperFinder DB=/work/projects/nn9305k/src/SeroTypeFinder/serotypefinder_db/ -cd /work/projects/nn9305k/src/SeroTypeFinder/ -python serotypefinder.py --help +python /work/projects/nn9305k/src/SeroTypeFinder/serotypefinder.py -i input_file -o output -d $DB conda deactivate ``` From 2b5ff6b378ab3e72b0f6fc27ecef06937af4b76c Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 11:23:21 +0200 Subject: [PATCH 53/83] Update ShigaTyper.md --- ShigaTyper.md | 40 +++++++++++++++++++++++++++------------- 1 file changed, 27 insertions(+), 13 deletions(-) diff --git a/ShigaTyper.md b/ShigaTyper.md index 5ae6e7d..f2854f3 100644 --- a/ShigaTyper.md +++ b/ShigaTyper.md @@ -1,20 +1,34 @@ -ShighaTyper -=========== -Original paper -"In Silico Serotyping Based on Whole-Genome Sequencing Improves the Accuracy of Shigella Identification." +**ShighaTyper** +--------------- +ShigaTyper is a quick and easy tool designed to determine Shigella serotype using Illumina paired end reads with low computation requirement. -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). -Location of Shigatyper -``` -/work/projects/nn9305k/src/ShigaTyper/shigatyper -``` +#### User Manual +https://bitbucket.org/genomicepidemiology/plasmidfinder/src + +#### For further reading +https://aem.asm.org/content/85/7/e00165-19 + +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory -Executing ShigaTyper -==================== ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=01:00:00 +#SBATCH --mem-per-cpu=32G + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + conda activate ShigaTyper -python /work/projects/nn9305k/src/ShigaTyper/shigatyper/shigatyper.py +python /work/projects/nn9305k/src/ShigaTyper/shigatyper/shigatyper.py Read1 Read2 -n sample_name conda deactivate ``` From 6b13e479e1b23b64787d01ff5272e654b42fd94b Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 11:27:16 +0200 Subject: [PATCH 54/83] Update VirulanceFinder.md --- VirulanceFinder.md | 30 ++++++++++++++++++++++++------ 1 file changed, 24 insertions(+), 6 deletions(-) diff --git a/VirulanceFinder.md b/VirulanceFinder.md index 4a74e57..d659889 100644 --- a/VirulanceFinder.md +++ b/VirulanceFinder.md @@ -1,19 +1,37 @@ **Executing VirulanceFinder** ----------------------------- -Don't need to mention about BLAST. Contact Jeevan if there is any issue. +VirulenceFinder identifies viruelnce genes in total or partial sequenced isolates of bacteria - at the moment only E. coli, Enterococcus, S. aureus and Listeria are available. +Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). -**The following code should be used inside a SLURM script.** -**We are preparing a full help page on how to run using SLURM script** +#### User Manual +https://bitbucket.org/genomicepidemiology/virulencefinder +#### For further reading +https://www.ncbi.nlm.nih.gov/pubmed/24574290 + +#### Here is the EXAMPLE SLURM script for Abel to excute the tool. +Important rules to follow +* Keep your data in /project/nn9305k/ +* Store your resutls also in /project/nn9305k/ +* Execute the script from your home directory ``` +#!/bin/bash +#SBATCH --job-name=DontKillMe +#SBATCH --account=nn9305k +#SBATCH --time=01:00:00 +#SBATCH --mem-per-cpu=32G + +## Set up job environment: +source /cluster/bin/jobsetup + +module load Miniconda3/4.4.10 + +# Activate Conda environment DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" conda activate VirulanceFinder - python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -p $DB - conda deactivate - ``` From bbc088b193291948744e6e7fc0551ad1204188eb Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 11:27:59 +0200 Subject: [PATCH 55/83] Update VirulanceFinder.md --- VirulanceFinder.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/VirulanceFinder.md b/VirulanceFinder.md index d659889..5efed80 100644 --- a/VirulanceFinder.md +++ b/VirulanceFinder.md @@ -29,9 +29,8 @@ source /cluster/bin/jobsetup module load Miniconda3/4.4.10 # Activate Conda environment -DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" - conda activate VirulanceFinder +DB="/work/projects/nn9305k/src/VirulanceFinder/Virulance_DB/virulencefinder_db" python /work/projects/nn9305k/src/VirulanceFinder/src/virulencefinder.py -p $DB conda deactivate ``` From 440f74ccf92779bd1ef4fbf817843a1f9e5c0f63 Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Fri, 5 Apr 2019 13:26:23 +0200 Subject: [PATCH 56/83] added the modified instruction for md5check --- working_with_hpc.md | 53 +++++++++++++++++++++++++++++++++++---------- 1 file changed, 41 insertions(+), 12 deletions(-) diff --git a/working_with_hpc.md b/working_with_hpc.md index ba64ac0..018d1b9 100644 --- a/working_with_hpc.md +++ b/working_with_hpc.md @@ -187,25 +187,54 @@ syntax for rsync? ### checking that file transfer completed without incident: You can either: -> - redo rsyn with same options: if the transfer was successfull, nothing will + - redo `rsync` with same options: if the transfer was successfull, nothing will be synchronized (same content). -> - use **hash** programs that generates a code based on file content + - use **hash** programs that generates a code based on file content (for both original and transfered/copied file). If both codes are identical, this means that the content of each files are identical i.e. that the file transfer was successull. -> use for ex. `md5sum file_origin`and `md5sum file_transfered`. The hash-codes +Example: `md5sum file_origin`and `md5sum file_transfered`. The hash-codes should be identical. -> Better to [automate] the process of checkingif you have many files -> - create a temporary file `tempfile_md5sum.txt` -> - generate md5sum for each files (use expressions) and append result to -`tempfile_md5sum.txt` -> - `md5sum yourfiles | tee "tempfile_md5sum.txt"` -> - `md5sum -c "tempfile_md5sum.txt"` -> - check the content of your file: it will idicate if there are differences -(if so, start again transfer for files where checksum does not match). -> - delete your temporary file `tempfile_md5sum.txt` +### For several files in the same folder: + +Better to automate the process if you have many files in the same folder + - create a temporary file `tempfile_md5sum.txt` + - generate md5sum for each files (use regular expressions) and append result to +`tempfile_md5sum.txt` Example: `md5sum yourfiles | tee "tempfile_md5sum.txt"` + - check the content of your file: it will idicate if there are differences: `md5sum -c "tempfile_md5sum.txt"` +(if there are differences, start again transfer for files where checksum did not match). + - delete your temporary file + +### For transfering all files within directories to a new location: + +Use rsync to copy all your files from directory to another (ie. moving your directories on Abel) + +Example: from `/work/projects/nn9305k/Mydirectory`to Mydirectory in parent_directory `/projects/nn9305k/` + +NB: **parent directory** (just the level above of where your want Mydirectory to be) + + 1) use `rsync -rauPW /origin_path/Mydirectory /parent_directory_destination` + + - NB: do not write /parent_directory_destination/Mydirectory otherwise it will make a subdirectory /../Mydirectory/Mydirectory + - Do it twice: if you get errors, it is probably a permissions problem -> contact Karin + +2) To compare if the copy/synchronisation of files from the origin and destination folders worked: we build 2 temporary files (containing the hash-codes for all files in origin and destination folders respectively). + +- You need to do it is from the respective parent directories of the origin and destination folders + +- the command finds all the files in your directories, executes md5sum on each file, sort hashes and then write into a file all those hash-codes. The idea is that if the copy/synchronisation succeded: then all the hashes codes for all files should be identical in the origin and destination directories. + +In the parent directory of origin : `find origin_Mydirectory -type f -exec md5sum {} + | sort -k 2 > origin_temp.txt` + +In the parent directory of destination: `find destination_Mydirectory -type f -exec md5sum {} + | sort -k 2 > destination_temp.txt` + +3) Then we check if the hash-codes generated for all the files in origin and destination directories are identical. It should return that files are identical. **If not, the file content of your directories are different** OR you did not do the previous command from the parent folder. + +`diff -s origin_mydirectory_temp.txt destination_mydirectory_temp.txt` + +4) When you are sure that the content of both directories are identical, you can remove the directory of origin, and the temporary files you created. ### wget From 3682d9e991e1341fef5b8ec64eabcefb3cbdaa35 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:11:18 +0200 Subject: [PATCH 57/83] Update SeroTypeFinder.md --- SeroTypeFinder.md | 4 +--- 1 file changed, 1 insertion(+), 3 deletions(-) diff --git a/SeroTypeFinder.md b/SeroTypeFinder.md index 18e000a..cc18de9 100644 --- a/SeroTypeFinder.md +++ b/SeroTypeFinder.md @@ -24,10 +24,8 @@ Important rules to follow ## Set up job environment: source /cluster/bin/jobsetup -module load Miniconda3/4.4.10 - conda activate SeroTyperFinder DB=/work/projects/nn9305k/src/SeroTypeFinder/serotypefinder_db/ -python /work/projects/nn9305k/src/SeroTypeFinder/serotypefinder.py -i input_file -o output -d $DB +python /work/projects/nn9305k/src/SeroTypeFinder/serotypefinder.py -i input_file -o output -p $DB -mp /work/projects/nn9305k/src/kma/kma conda deactivate ``` From 5da7dbfcd3f14df3beab016afae50cde8fb382cc Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:12:06 +0200 Subject: [PATCH 58/83] Update PlasmidFinder.md --- PlasmidFinder.md | 2 -- 1 file changed, 2 deletions(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 7f66790..521646f 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -26,8 +26,6 @@ Important rules to follow ## Set up job environment: source /cluster/bin/jobsetup -module load Miniconda3/4.4.10 - # Activate Conda environment conda activate PlasmidFinder From e95f0d1ce97e547d03255782244b8f34a4936ec6 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:19:57 +0200 Subject: [PATCH 59/83] Update PlasmidFinder.md --- PlasmidFinder.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/PlasmidFinder.md b/PlasmidFinder.md index 521646f..f8e69cc 100644 --- a/PlasmidFinder.md +++ b/PlasmidFinder.md @@ -1,7 +1,6 @@ **PlasmidFinder** ------------------------- PlasmidFinder identifies plasmids in total or partial sequenced isolates of bacteria. - Contact Jeevan in slack if you have any issues or further assistance (F. ex. run the tool for multiple isolates). #### User Manual @@ -12,6 +11,7 @@ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4068535/ #### Here is the EXAMPLE SLURM script for Abel to excute the tool. Important rules to follow +* Refer the user manual for all the parameters in the tool * Keep your data in /project/nn9305k/ * Store your resutls also in /project/nn9305k/ * Execute the script from your home directory From e97f16785cf9f1453acb018f4622cace29531392 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:20:35 +0200 Subject: [PATCH 60/83] Update PointFinder.md --- PointFinder.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/PointFinder.md b/PointFinder.md index 18f0bb1..b9a642e 100644 --- a/PointFinder.md +++ b/PointFinder.md @@ -9,6 +9,7 @@ https://bitbucket.org/genomicepidemiology/pointfinder #### Here is the EXAMPLE SLURM script for Abel to excute the tool. Important rules to follow +* Refer the user manual for all the parameters in the tool * Keep your data in /project/nn9305k/ * Store your resutls also in /project/nn9305k/ * Execute the script from your home directory @@ -23,8 +24,6 @@ Important rules to follow ## Set up job environment: source /cluster/bin/jobsetup -module load Miniconda3/4.4.10 - conda activate PointFinder # Database location From 1ede76774022c57a70761db99d5803a37a99c33e Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:21:12 +0200 Subject: [PATCH 61/83] Update ResFinder.md --- ResFinder.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/ResFinder.md b/ResFinder.md index a388a20..553bc1a 100644 --- a/ResFinder.md +++ b/ResFinder.md @@ -12,6 +12,7 @@ https://bitbucket.org/genomicepidemiology/pointfinder #### Here is the EXAMPLE SLURM script for Abel to excute the tool. Important rules to follow +* Refer the user manual for all the parameters in the tool * Keep your data in /project/nn9305k/ * Store your resutls also in /project/nn9305k/ * Execute the script from your home directory @@ -26,8 +27,6 @@ Important rules to follow ## Set up job environment: source /cluster/bin/jobsetup -module load Miniconda3/4.4.10 - conda activate ResFinder # Location of PointFinder DB From fb47266b1478d738ec189ae4a2e2efc83d796b16 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:23:54 +0200 Subject: [PATCH 62/83] Update SeqSero.md --- SeqSero.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/SeqSero.md b/SeqSero.md index 3b73740..b8499ed 100644 --- a/SeqSero.md +++ b/SeqSero.md @@ -12,6 +12,7 @@ http://jcm.asm.org/content/early/2015/03/05/JCM.00323-15 #### Here is the EXAMPLE SLURM script for Abel to excute the tool. Important rules to follow +* Refer the user manual for all the parameters in the tool * Keep your data in /project/nn9305k/ * Store your resutls also in /project/nn9305k/ * Execute the script from your home directory @@ -26,8 +27,6 @@ Important rules to follow ## Set up job environment: source /cluster/bin/jobsetup -module load Miniconda3/4.4.10 - conda activate SeqSero_Shared python /work/projects/nn9305k/src/SeqSero/SeqSero/SeqSero.py -m "1" -i input_file -b "mem" conda deactivate From 7cbbb55d2e2e10811d04f9abffe33df9ed4ca700 Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:24:28 +0200 Subject: [PATCH 63/83] Update SeroTypeFinder.md --- SeroTypeFinder.md | 1 + 1 file changed, 1 insertion(+) diff --git a/SeroTypeFinder.md b/SeroTypeFinder.md index cc18de9..9d7f32c 100644 --- a/SeroTypeFinder.md +++ b/SeroTypeFinder.md @@ -10,6 +10,7 @@ https://bitbucket.org/genomicepidemiology/serotypefinder/src/master/ #### Here is the EXAMPLE SLURM script for Abel to excute the tool. Important rules to follow +* Refer the user manual for all the parameters in the tool * Keep your data in /project/nn9305k/ * Store your resutls also in /project/nn9305k/ * Execute the script from your home directory From 7bb9c59d4cbf7dee502f0063b10e72790e4c576a Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Fri, 5 Apr 2019 16:24:40 +0200 Subject: [PATCH 64/83] updated poppunk - backup --- poppunk_tree.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/poppunk_tree.md b/poppunk_tree.md index 228fd77..28cabc6 100644 --- a/poppunk_tree.md +++ b/poppunk_tree.md @@ -1,4 +1,9 @@ # 1. What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) : Population Partitioning Using Nucleotide K-mers + +[ubioinfo virtual lab talk 2019-04-01 John Lees](https://youtu.be/uEoah6_XSSE) + +- less good for low diversity species, and when blur (lots of recombination) +- ## 1.1 Summary - Overview [Manual](https://poppunk.readthedocs.io/) From e0f877c555f9df1591d483068f3583d1c72976ab Mon Sep 17 00:00:00 2001 From: Jeevan Karloss Date: Fri, 5 Apr 2019 16:25:02 +0200 Subject: [PATCH 65/83] Update ShigaTyper.md --- ShigaTyper.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/ShigaTyper.md b/ShigaTyper.md index f2854f3..b74b7b6 100644 --- a/ShigaTyper.md +++ b/ShigaTyper.md @@ -12,6 +12,7 @@ https://aem.asm.org/content/85/7/e00165-19 #### Here is the EXAMPLE SLURM script for Abel to excute the tool. Important rules to follow +* Refer the user manual for all the parameters in the tool * Keep your data in /project/nn9305k/ * Store your resutls also in /project/nn9305k/ * Execute the script from your home directory @@ -26,8 +27,6 @@ Important rules to follow ## Set up job environment: source /cluster/bin/jobsetup -module load Miniconda3/4.4.10 - conda activate ShigaTyper python /work/projects/nn9305k/src/ShigaTyper/shigatyper/shigatyper.py Read1 Read2 -n sample_name conda deactivate From 49ce53315d5f0f35ee2265df12b257ad421d16a9 Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Sat, 6 Apr 2019 13:42:45 +0200 Subject: [PATCH 66/83] added workflow fig --- poppunk_tree.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/poppunk_tree.md b/poppunk_tree.md index 28cabc6..d54f744 100644 --- a/poppunk_tree.md +++ b/poppunk_tree.md @@ -20,6 +20,8 @@ - # 2. How to use PopPUNK + + ## 2.1 Create datadase - create a list of your assemblies and other sequences you want to include: you need to have the path of the file included, so you need to create the list from the folder you will run PopPUNK: `ls /*fasta > reference_list.txt` From 2bd364d584a08519342426f3702c0db047771393 Mon Sep 17 00:00:00 2001 From: evezeyl Date: Sat, 6 Apr 2019 19:46:22 +0200 Subject: [PATCH 67/83] on the way to be clear --- poppunk_tree.md | 228 +++++++++++++++++++++++------------------------- 1 file changed, 110 insertions(+), 118 deletions(-) diff --git a/poppunk_tree.md b/poppunk_tree.md index d54f744..11109ea 100644 --- a/poppunk_tree.md +++ b/poppunk_tree.md @@ -1,6 +1,8 @@ # 1. What is [PopPUNK](https://genome.cshlp.org/content/29/2/304) : Population Partitioning Using Nucleotide K-mers -[ubioinfo virtual lab talk 2019-04-01 John Lees](https://youtu.be/uEoah6_XSSE) +Presentation video: [ubioinfo virtual lab talk 2019-04-01 John Lees](https://youtu.be/uEoah6_XSSE) + +[Methods and interpretation presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) - less good for low diversity species, and when blur (lots of recombination) - @@ -10,7 +12,7 @@ [PopPUNK Documentation](https://poppunk.readthedocs.io/en/latest/index.html) [Presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) -- whole genome (core + assessory) population analysis/clustering +- whole genome (core + accessory) population analysis/clustering - distinction between isolates : uses k-mer of different length: `mash` to find core and accessory distances between isolates (**pairwise**) - the distribution of those distances is used to discriminate clusters (defined as strains) of closely related isolates (similarity:both core and accessory) - clustering of newly added isolates: EXTENDABLE4 - without the need of reanalizing all samples @@ -18,103 +20,149 @@ > aimed for consistent naming clusters between studies, outbreak detection in minutes - analyse up to 10^4 samples in single step - possible to add new samples - -# 2. How to use PopPUNK - +# 2. How to use PopPUNK: first steps ## 2.1 Create datadase -- create a list of your assemblies and other sequences you want to include: you need to have the path of the file included, so you need to create the list from the folder you will run PopPUNK: `ls /*fasta > reference_list.txt` -- -- +- create a list of your assemblies and other sequences you want to include in the analysis: +`NB`: You need to have the path of the file included, so you need to create the list from the folder you will run PopPUNK: +`ls > reference_files.txt` -poppunk --fit-model --distances --ref-db --output --full-db --K +- There are 2 solutions to create the database: -## 2.2 Fitting a model: what to look for +```bash +# only database +poppunk --create-db --r-files --output \ + --threads <2> --plot-fit <5> +# OR +# database and first model fitting in one step (see bellow) +poppunk --easy-run --r-files --output --threads <4> --plot-fit <5> --min-k <13> --full-db +``` +-------------------------------------------------------------------------------- +What happens: +1) reference files are hashed at different k-mer lenghts (using [mash](https://mash.readthedocs.io/en/latest/index.html). +2) pairwise distances between sequences are calculated +> Accessory and core distances are estimated jointly: through probabilities estimates of the number of k-mers matching over a range of k-mer sizes. Decomposing accessory and core distance is allowed because: +- Small k-mer size allow accessory distance to be independent of k-mer size (but k-mers size must be large enough such as k-mers do not match randomly between sequences) +- EXPLain better +core distances are estimated through the range of k-mer size (estimate density SNPs over ) -

- -

+#picture +-------------------------------------------------------------------------------- +`inspect the plots created and the console output`, to determine if the range of k-mer size used is adequate: +- #kmer must be inferior <- stats here +- #the plots represent: regressions of log(pr(pa,b): the proportion of k-mers matching at length k between sequences a and b (for random sample pairs a and b): -> trick for seing plot : test! - [ ] use move with bondary with 0 +Read [here](https://poppunk.readthedocs.io/en/latest/troubleshooting.html#kmer-length) if there is adjusting need of the k-mer size-range. Ex: add options `--min-k <15> --kmer-step <2>` + +## 2.2 Fitting a model + +![fitting](./figures/refine_poppunk.png) + +Blobs: will represent distances between hierarchies of distances (ex: within strains, between strains, between populations, clones...) + +You need to choose between 2D Gaussian mixed model () OR DBSCAN models to fit the distribution of core and accessory distance data: +- `--K ` (default blobs = 2): +> equal likelihood contours and decision boundary (within and between cluster-assignments ) +- `--dbscan` will estimate the number of blobs (representing distances between different hierarchies) for you -> kernet density estimate - > can be used to identify outliers and contamination (program to remove those isolates from DB) > - [ ] where? +`poppunk --fit-model --distances --ref-db \ +--output --full-db` + a model option ex: `--K ` +`--model` will also copy over the model fit, so that the entire PopPUNK database is in a single directory +`--external-clustering` one can provide cluster names or labels that have been previously defined by any other method. This could include, for example, another clustering methods IDs, serotypes, clonal complexes and MLST assignments. The input is a CSV file which is formatted as follows: + +sample,serotype,MLST +sample1,12,34 +sample2,23F,1 + +> trick for seing plot : test! - [ ] use move with bondary with 0 + +> kernel density estimate - > can be used to identify outliers and contamination (program to remove those isolates from DB) > - [ ] where? - how to do: accessory, core, and combined clusters (see sup material) - [ ] -### 2.2.1 2D GMM -> equal likelihood contours and decision boundary (within and between cluster-assignments ) +### Evaluate the model: + +- Model: Fit summary: + -- `Number of components used`: represents the number of blobs (--K) <=> the number of hierarchies + -- `Avg. entropy of assignment`: confidence of assignment to blobs (near 0 = well defined = confidence) -### 2.2.2 HDBSCAN - -`--dbscan` +- Network summary: + -- components used <=> the number of populations/clusters (not blobs) + -- Transitivity: must be high (near 1 -> everything within a cluster belongs to same strain/cluster-groups <=> fewer within- than between-strain links) + -- network density **must be low** (means few isolate per group <=> good separation between population/cluster-groups: meaning fewer within than between strains links + -- `network score` = **summary for evaluating model** -> must be close to 1 (>0.8). Score is calculated using density and transitivity -### 2.2.3 Evaluate the chosen model -- silhouette index = measure of how similar an object is to its own cluster (cohesion) compared to other clusters (separation) -> must be close to 1 -- network density must be low (mean good separation between clusters: fewer within than between strains links) +In the network: edges are pairwise distances, nodes are isolates + +[More technical explanations:](https://poppunk.readthedocs.io/en/latest/tutorial.html#creating-a-database) + +If your model is not satisfactory +- blobs not well delimited: you can try to increase sketch sizes when creating distances data-basis +- adjust your k-mer size range +- adjust boundaries (how core and accessory distances are used to discriminate between clusters) > **important to look at the network** +#Not that+ - `silhouette index`?? = measure of how similar an object is to its own cluster (cohesion) compared to other clusters (separation) -#### 2.2.4 Only if the model is really bad: (otherwise -> go to refine model) -- possibility to optimize using core distance only (vertical boundary) or accessory only (horizontal boundary) -> when? if core and accessory genomes have independent evolution histories - ex. lots of recombination (blurs in blobs), prophages... -> NB: depending on level problem - can be sufficient to only refine model +#### 2.3 Model refinement (improvement model) `--refine-model` -- **modify sketch size** (increase sensibility detection SNPs also increases running time) -> finer differences in %pi (down to single SNPs) --option -[ ] + Tweak the existing fit and pick a better boundary to distinguish within and between strains. -## 2.3. refining the model `--refine-model` +`poppunk --refine-model --distances --model-dir --ref-db distances_db --full-db --output --full-db ` -`poppunk --refine-model --distances --ref-db --output --full-db` + For 2D GMM `--K ` OR For HDBSCAN `--dbscan` +Model options: +- For 2D GMM `--K ` +- For HDBSCAN `--dbscan` -Common optional options: HDBSCAN & 2D GMM +- Boundary shifting options `--pos-shift ` (away from origin) `--neg-shift ` (towards origin) -`--manual-start ` -`--indiv-refine` allow these boundaries to be placed independently on core/accessory -

-> need to create a triangular⁵ boundary - move forward and backward FROM starting point (range) -``` -mean0 (x,y) #for within strains blob -mean1 (x,y) #for between strains blob -start (x,y) #starting point to move boundary to -``` -

+> Re-evaluate your model +-------------------------------------------------------------------------------- +For difficult data sets: -example: `poppunk --refine-model --distances lm_example/lm_example.dists --ref-db lm_example_2DGMM --output refine_gmm --full-db --neg-shift 0.1` -to obtain the same refinement as published in their article +The models assume that core and accessory distances are correlated. This might not be true for: +- species/groups showing "low diversity" -> a and %pi not necessary correlated - analyze independently (see [supplement article](https://www.biorxiv.org/content/10.1101/360917v2.supplementary-material?versioned=true)) +- it is also possible to optimize models using core distance only (vertical boundary) or accessory only (horizontal boundary). Useful when/if core and accessory genomes have independent evolution histories. (ex. lots of recombination, blurs in blobs, lots insertions eg. prophages...) + `--indiv-refine` allow these boundaries to be placed independently on core/accessory + `--manual-start ` + > need to create a triangular⁵ boundary - move forward and backward FROM starting point (range) + ``` + mean0 (x,y) #for within strains blob + mean1 (x,y) #for between strains blob + start (x,y) #starting point to move boundary to + ``` -NB: **Better to redo all the steps in the same folder - now that we now the best fit - to facilitate further use** +**Modifing sketch size when creating distance database** (increase sensibility detection SNPs also increases running time) -> but allow a better discrimination between distances. Theoretically down to single SNPs ## 2.4. Simplification database (optional) -When model is good -> then we can stop using `--full-db` option (but not compulsory) - -## 2.5 Visualisation resuts -> output can be made either from --model-fit or --refine-model -`--microreact` Generate output files for microreact visualisation -`--cytoscape `Generate network output files for Cytoscape -`--phandango` Generate phylogeny and TSV for Phandango visualisation -`--grapetree` Generate phylogeny and CSV for grapetree visualisation -`--rapidnj` RAPIDNJ (Path to rapidNJ binary to build NJ tree for Microreact) -`--perplexity `PERPLEXITY (Perplexity used to calculate t-SNE projection (with –microreact) [default=20.0]) -`--info-csv` INFO_CSV (additional metadata: Epidemiological information CSV formatted for microreact (can be used with other outputs)) +When model is good -> then we can stop using `--full-db` option (but not compulsory). See manual. +## 2.5 Visualization resuts +Output can be made either from `--model-fit` or `--refine-model`, using the same parameters that lead to a good model with `-- use-model` -### 2.5.1 Microreact +`--microreact` Generate output files for [Microreact] visualisation -### 2.5.2.GrapeTree +`--rapidnj` RAPIDNJ (Path to rapidNJ software if you want to build rapidNJ tree for [Microreact]) +`--info-csv` INFO_CSV (additional metadata: Epidemiological information CSV formatted for microreact (can be used with other outputs)) -### 2.5.3. Phandango +`--cytoscape `Generate network output files for [Cytoscape] +`--phandango` Generate phylogeny and TSV for [Phandango] visualisation +`--grapetree` Generate phylogeny and CSV for [Grapetree] visualisation -### 2.5.4 Cytoscape +`--perplexity `PERPLEXITY (Perplexity used to calculate t-SNE projection (with –microreact) [default=20.0]) ## 2.6 Adding new sequences = assigning queries @@ -135,70 +183,14 @@ When model is good -> then we can stop using `--full-db` option (but not compuls --ref-db `--core-only` --ref-db `--accessory-only ` +________________________________________________________________________________ +[Microreact]:https://microreact.org/showcase +[Cytoscape]: +[Phandango]: +[Grapetree] XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX - - - - -# Poppunk commands - - -# Trics for difficult species: -- Low diversity -> a and %pi not necessary correlated - analyse independently (see supplement) - - genome wide association study: genes responsible differences in a and %pi clusters (pyseer) -> used a-cluster as phenotype - - -# My Points -- [ ] as use random sampling of distances to create network -> maybe important not to unbalanced sampling to create model -> otherwise model risicate to be pushed toward most common lineage (specially if expect new samples different) - same pb as usual ... -- [ ] how to extracti within and between-cluster SNPs distances -> %pi?? -- [ ] understand this sketch size -> per scaffold or over whole genome - must be high!! -> the scetch size -> determines what will be most computationnaly intensive - -- within cluster distance: appear under-estimate - but between comparable to RhierBAPS (maybe depend on model used) - -# ## 2.opening PopPUNK (how does it functions) - -- p_random : maximum probability of random matches kmerse (set to 5% ?? check) -> determines k_min -- kmax set at 29 by default (can increase to max mash is able to support = - [ ] find ) - -## 2.1 Models: classify which distances pairs (%pi and a) are within the same cluster - -### 2.1.1 Model to fit -> Two-dimensional Gaussian mixture model (2D GMM) -Fits random subsample of up to 10⁵ distance pairs -scikit-learn 0.19 -Dirichlet Process prior on weights (nearly 0 to define within-cluster distances) -best final likelihood from five k-means initial starts -K: maximum allowed of mixture components (default = 2) -Distances classified with fitted model - -### 2.1.2 Model to fit -> HDBSCAN (option dbscan) -Classify a subsample of 10⁵ points (distance pairs) using: -Boruvka ball tree Algorithm -Iterations - progressive reductions -> minimum of samples required to initiate the seqrch for a cluster - - -> how conservative clustering is - - -> minimum cluster size (threshold nb points(distances a cluster must contain) - - -> extends points in cluster closest to origin (represent within strains) - should not overlap with between strains clusters - -### 2.2 Networks constructed: -networkx v2.1 -> undirected graph with unweighted egdes -> population clusters -extraction: -- connected components (nodes = samples, connections = pairwise distances) -- ordered by nb isolates (largest to smallest) -Evaluation netwok structure: ns = transitivity (1 - density) -- [ ] transitivity? -- (1-density) -> to subdivise the population - -ns > 0.8 - 1 good fit - -database simplification: randomly select one isolate - per cluster (contected to all other members clusters) - -### 2.3 Refinement distance classification -Models -> treat clusters symmetrically (force clusters same distance/structure) -Refinement -> delimit precisely range %pi - a distances trated as within strain links - to maximize ns - - line between means of within and between strain clusters - - if not correct fit -> provide within and between stran cluster means manually - move boundary starting point (range of move) -> maximize ns (40 equaly spaced points over the allowed range) - # Summary - Files produced | File extension | what | when | From 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0 HcmV?d00001 diff --git a/poppunk_tree.md b/poppunk_tree.md index 11109ea..4a31a97 100644 --- a/poppunk_tree.md +++ b/poppunk_tree.md @@ -2,25 +2,30 @@ Presentation video: [ubioinfo virtual lab talk 2019-04-01 John Lees](https://youtu.be/uEoah6_XSSE) -[Methods and interpretation presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) +[Introduction to Methods and interpretation: presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) -- less good for low diversity species, and when blur (lots of recombination) -- ## 1.1 Summary - Overview -[Manual](https://poppunk.readthedocs.io/) -[PopPUNK Documentation](https://poppunk.readthedocs.io/en/latest/index.html) -[Presentation](https://docs.google.com/presentation/d/1StmmM02lSpFPdevQT3iDB3BAKRoMC8Q4NtJ4nzu7MdY/edit?usp=sharing) - -- whole genome (core + accessory) population analysis/clustering -- distinction between isolates : uses k-mer of different length: `mash` to find core and accessory distances between isolates (**pairwise**) -- the distribution of those distances is used to discriminate clusters (defined as strains) of closely related isolates (similarity:both core and accessory) -- clustering of newly added isolates: EXTENDABLE4 - without the need of reanalizing all samples -- maintenance free and auto-reduce database -> aimed for consistent naming clusters between studies, outbreak detection in minutes -- analyse up to 10^4 samples in single step - possible to add new samples -- +- whole genome (core + accessory) population analysis/clustering. Coding and non-coding. +- distinction between isolates : uses `mash` with a range of k-mers at different lengths to estimate **pairwise core and accessory distances** between isolates +- the distribution of pairwise distances is used to discriminate between clusters of isolates: within or between strains (and discrimination of different hierarchical level of distances). +- When the model used for clustering has been defined: new isolates can easily be added, without the need to re-analyze all samples. Clusters naming remain consistent, and metadata import allows using previously defined naming (ie. MLST) +- Database maintenance free, the size of the database can be kept small by using representative isolates for each cluster (automatically chosen) once the model has been fitted for your species of interest. +- predefined database with model fit can be found at: +- This software has been developed for rapid outbreak detection. It is possible to analyze up to 10^4 samples in a single steps. + # 2. How to use PopPUNK: first steps + +[Manual](https://poppunk.readthedocs.io/). Note that the manual has been written for slightly different versions of PopPUNK. Therefore some command names in the documentation might be slightly different than the PopPUNK version installed on Abel. + +Activate PopPUNK: `source activate poppunk` + +To check command synthax: `poppunk --help` + +PopPUNK workflow is really simple, but can appear complicated because several options can be combined and several steps can be done at once. Keep it simple: do one step at the time to understand what it does. + +Commands that can be combined: [Overview](https://poppunk.readthedocs.io/en/latest/options.html) + ## 2.1 Create datadase @@ -31,33 +36,44 @@ Presentation video: [ubioinfo virtual lab talk 2019-04-01 John Lees](https://you `ls > reference_files.txt` - There are 2 solutions to create the database: +1) the quick start option: `--easy-run` does 2 steps at once (not recommended for learning) which does both create the database and does the first model fitting +2) Use the `--create-db` option which only creates: the _mash sketches database_ and the _distances database_ (also create a file containing isolate names) ```bash # only database poppunk --create-db --r-files --output \ --threads <2> --plot-fit <5> - -# OR -# database and first model fitting in one step (see bellow) -poppunk --easy-run --r-files --output --threads <4> --plot-fit <5> --min-k <13> --full-db ``` -------------------------------------------------------------------------------- -What happens: -1) reference files are hashed at different k-mer lenghts (using [mash](https://mash.readthedocs.io/en/latest/index.html). +What is happening: +1) reference files are hashed at different k-mer lenghts using [mash](https://mash.readthedocs.io/en/latest/index.html). 2) pairwise distances between sequences are calculated -> Accessory and core distances are estimated jointly: through probabilities estimates of the number of k-mers matching over a range of k-mer sizes. Decomposing accessory and core distance is allowed because: -- Small k-mer size allow accessory distance to be independent of k-mer size (but k-mers size must be large enough such as k-mers do not match randomly between sequences) -- EXPLain better -core distances are estimated through the range of k-mer size (estimate density SNPs over ) +Accessory and core distances are estimated jointly: through probabilities estimates of the number of k-mers matching over a range of 5 k-mer sizes. (5 k-mer sizes has been evaluated to provide a good estimate) +Decomposing accessory and core distance is possible because: small k-mer size allow accessory distance to be independent of k-mer size (but k-mers size must be large enough such as k-mers do not match randomly between sequences) #picture -------------------------------------------------------------------------------- -`inspect the plots created and the console output`, to determine if the range of k-mer size used is adequate: -- #kmer must be inferior <- stats here -- #the plots represent: regressions of log(pr(pa,b): the proportion of k-mers matching at length k between sequences a and b (for random sample pairs a and b): +**Evaluate:** inspect the plots created and the console output, to determine if the range of k-mer size used is adequate: +``` +Creating mash database for k = 13 +Random 13-mer probability: 0.04 +Found existing mash database distances_db/distances_db.13.msh for k = 13 +Creating mash database for k = 17 +Random 17-mer probability: 0.00 +... +``` +- Random k-mer probability match must be inferior to 5% - if you want to be more stringent, you can increase the minimum k-mer length: `--min-k <17>`. Default step between k-mer sizes is set to 4. You can also change that using: `--k-step <4>` Read [here](https://poppunk.readthedocs.io/en/latest/troubleshooting.html#kmer-length) if necessary. NB: if k-mer size is to big you will get a warning: `outliers` + +- the plots represent: regressions of log(pr(pa,b): the proportion of k-mers matching at length k between sequences a and b (for random sample pairs a and b). You want a straight line, and no outliers + +

+ +

+ +If the assembly length of your isolates is highly variable (due to structural variation) you can add the option `--ignore-length` which will ignore outliers due to sequence length. -Read [here](https://poppunk.readthedocs.io/en/latest/troubleshooting.html#kmer-length) if there is adjusting need of the k-mer size-range. Ex: add options `--min-k <15> --kmer-step <2>` +If your species has really low diversity you can increase sketch size. Here is default: `--sketch-size <10000>` ## 2.2 Fitting a model From 8dca419f9ee18634aef42b107d3682dfcab5c6f5 Mon Sep 17 00:00:00 2001 From: evezeyl <45561997+evezeyl@users.noreply.github.com> Date: Tue, 9 Apr 2019 16:58:17 +0200 Subject: [PATCH 69/83] almost there with poppunk --- figures/poppunk/2DGMM_fit_DPGMM_fit.png | Bin 0 -> 142996 bytes .../2DGMM_fit_distanceDistribution.png | Bin 0 -> 148831 bytes figures/poppunk/DBSCAN_refine_refined_fit.png | Bin 0 -> 178801 bytes figures/{ => poppunk}/poppunk_random_k.png | Bin figures/{ => poppunk}/refine_poppunk.png | Bin poppunk_tree.md | 317 ++++++++++++------ 6 files changed, 206 insertions(+), 111 deletions(-) create mode 100644 figures/poppunk/2DGMM_fit_DPGMM_fit.png create mode 100644 figures/poppunk/2DGMM_fit_distanceDistribution.png create mode 100644 figures/poppunk/DBSCAN_refine_refined_fit.png rename figures/{ => poppunk}/poppunk_random_k.png (100%) rename figures/{ => poppunk}/refine_poppunk.png (100%) diff --git 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