From f9dfc907cb3342a9b3a48c7b4dde25c3f5926936 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 1 Nov 2024 13:25:06 -0700 Subject: [PATCH 01/15] update figure and workflow overview --- docs/index.rst | 88 +++++++++++++++++++----------------------- docs/rqc_workflow.png | Bin 62533 -> 38172 bytes 2 files changed, 40 insertions(+), 48 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 73b5b93..b055f3c 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -9,35 +9,43 @@ Reads QC Workflow (v1.0.2) Workflow Overview ----------------- -This workflow utilizes the program “rqcfilter2” from BBTools to perform quality control on raw Illumina reads. The workflow performs quality trimming, artifact removal, linker trimming, adapter trimming, and spike-in removal (using BBDuk), and performs human/cat/dog/mouse/microbe removal (using BBMap). - -The following parameters are used for "rqcfilter2" in this workflow:: - - qtrim=r : Quality-trim from right ends before mapping. - - trimq=0 : Trim quality threshold. - - maxns=3 : Reads with more Ns than this will be discarded. - - maq=3 : Reads with average quality (before trimming) below this will be discarded. - - minlen=51 : Reads shorter than this after trimming will be discarded. Pairs will be discarded only if both are shorter. - - mlf=0.33 : Reads shorter than this fraction of original length after trimming will be discarded. - - phix=true : Remove reads containing phiX kmers. - - khist=true : Generate a kmer-frequency histogram of the output data. - - kapa=true : Remove and quantify kapa tag - - trimpolyg=5 : Trim reads that start or end with a G polymer at least this long - - clumpify=true : Run clumpify; all deduplication flags require this. - - removehuman=true : Remove human reads via mapping. - - removedog=true : Remove dog reads via mapping. - - removecat=true : Remove cat reads via mapping. - - removemouse=true : Remove mouse reads via mapping. - - barcodefilter=false : Disable improper barcodes filter - - chastityfilter=false: Remove illumina reads failing chastity filter. - - trimfragadapter=true: Trim all known Illumina adapter sequences, including TruSeq and Nextera. - - removemicrobes=true : Remove common contaminant microbial reads via mapping, and place them in a separate file. +This workflow utilizes the program "rqcfilter2" from BBTools to perform quality control on raw Illumina reads. The workflow performs quality trimming, artifact removal, linker trimming, adapter trimming, and spike-in removal (using BBDuk), and performs human/cat/dog/mouse/microbe removal (using BBMap). + +The following parameters are used for "rqcfilter2" in this workflow: + - barcodefilter=false : Disable improper barcodes filter + - chastityfilter=false: Remove illumina reads failing chastity filter. + - clumpify=true : Run clumpify; all deduplication flags require this. + - extend=false : Extend reads during merging to allow insert size estimation of non-overlapping reads. + - jni=true : Enable C code for higher speed and identical results. + - usejni=false : Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code; details are in /jni/README.txt. + - khist=true : Generate a kmer-frequency histogram of the output data. + - maq=10 : Reads with average quality (before trimming) below this will be discarded. + - maxns=1 : Reads with more Ns than this will be discarded. + - minlen=51 : Reads shorter than this after trimming will be discarded. Pairs will be discarded only if both are shorter. + - mlf=0.33 : Reads shorter than this fraction of original length after trimming will be discarded. + - mtst=true : Spike-in bbduk removal mtst parameter + - phix=true : Remove reads containing phiX kmers. + - pigz=true : Use pigz for compression + - qtrim=r : Quality-trim from right ends before mapping. + - removecat=true : Remove cat reads via mapping. + - removedog=true : Remove dog reads via mapping. + - removehuman=true : Remove human reads via mapping. + - removemicrobes=true : Remove common contaminant microbial reads via mapping, and place them in a separate file. + - removemouse=true : Remove mouse reads via mapping. + - removeribo=true : Remove ribosomal reads via kmer-matching, and place them in a separate file. + - **rna=true** : Parameter for RNA-seq analysis. + - sketch=true : Run SendSketch on 2M read pairs. + - trimfragadapter=true: Trim all known Illumina adapter sequences, including TruSeq and Nextera. + - trimq=0 : Trim quality threshold. + - trimpolyg=5 : Trim reads that start or end with a G polymer at least this long. + - unpigz=t : Use pigz for decompression Workflow Availability --------------------- The workflow from GitHub uses all the listed docker images to run all third-party tools. -The workflow is available in GitHub: https://github.com/microbiomedata/ReadsQC; the corresponding +The workflow is available in GitHub: https://github.com/microbiomedata/metaT_ReadsQC; the corresponding Docker image is available in DockerHub: https://hub.docker.com/r/microbiomedata/bbtools. Requirements for Execution @@ -63,6 +71,7 @@ Third party software (This is included in the Docker image.) - `BBTools v38.96 `_ (License: `BSD-3-Clause-LBNL `_) + Requisite database ~~~~~~~~~~~~~~~~~~ @@ -78,20 +87,8 @@ The following commands will download the database:: Sample dataset(s) ----------------- -- small dataset: `Ecoli 10x `_ . You can find input/output in the downloaded tar gz file. - -- large dataset: Zymobiomics mock-community DNA control (`SRR7877884 `_); the `original gzipped dataset `_ is ~5.7 GB. You can find input/output in the downloaded tar gz file. - - -.. note:: - - If the input data is paired-end data, it must be in interleaved format. The following command will interleave the files, using the above dataset as an example: - -.. code-block:: bash +- Bulk soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_122 (`SRR8552838 `_)` with `metadata available in the NMDC Data Portal `_ - paste <(zcat SRR7877884_1.fastq.gz | paste - - - -) <(zcat SRR7877884_2.fastq.gz | paste - - - -) | tr '\t' '\n' | gzip -c > SRR7877884-int.fastq.gz - -For testing purposes and for the following examples, we used a 10% sub-sampling of the above dataset: `SRR7877884-int-0.1.fastq.gz `_. This dataset is already interleaved. Inputs ------ @@ -113,18 +110,13 @@ An example input JSON file is shown below: .. code-block:: JSON { - "jgi_rqcfilter.database": "/path/to/refdata", - "jgi_rqcfilter.input_files": [ - "/path/to/SRR7877884-int-0.1.fastq.gz " - ], - "jgi_rqcfilter.input_interleaved": true, - "jgi_rqcfilter.input_fq1":[], - "jgi_rqcfilter.input_fq2":[], - "jgi_rqcfilter.outdir": "/path/to/rqcfiltered", - "jgi_rqcfilter.memory": "35G", - "jgi_rqcfilter.threads": "16" - } + "metaTReadsQC.input_files": ["/global/cfs/cdirs/m3408/test_data/metatranscriptome/antisense/52400.2.318150.GATACTGG-CCAGTATC.fastq.gz"], + "metaTReadsQC.proj":"nmdc:xxxxxxx", + "metaTReadsQC.rqc_mem": 180, + "metaTReadsQC.rqc_thr": 64, + "metaTReadsQC.database": "/refdata/" + } .. note:: In an HPC environment, parallel processing allows for processing multiple samples. The "jgi_rqcfilter.input_files" parameter is an array data structure. It can be used for multiple samples as input separated by a comma (,). @@ -162,7 +154,7 @@ Below is an example of all the output directory files with descriptions to the r ==================================== ============================================================================ FileName Description ==================================== ============================================================================ -SRR7877884-int-0.1.anqdpht.fastq.gz main output (clean data) +nmdc_xxxxxx.fastq.gz main output (clean data) adaptersDetected.fa adapters detected and removed bhist.txt base composition histogram by position cardinality.txt estimation of the number of unique kmers diff --git a/docs/rqc_workflow.png b/docs/rqc_workflow.png index 35c8cce59d38094418d155e96d8f6e91cf3cb3ac..44dbe29d52c0ce0d8c525cfddec533b48f04a6b6 100644 GIT binary patch literal 38172 zcmZU*by$_p^FDqk>5!5RNhPHl1eES>kd*H3R78<(kW{+6Q9-)9K|nw_)S=_M$JhJw z`{(C%ad~(=&+hE*%suza?D$?)Sq2+}3Jt&7yqYLARZR{>kfujOz^8dUB#EUU 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zTnY?l`g&Q>ykt+08ZzkUr(S*SN)P0@F)*z1iCRT>E=0gn0fs<(s)l|6k_imE43ll? zXnOz)1@@_Bnr#`~DINh=0fzf3;#E=xIeVha=3cj{0WN g15y3|OYj}eSspHl#jzUX0pRzCzKLFy?v-2r1I&vwR{#J2 From 588ef7e4033f983ca2462156e7602c436a50b29f Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Sun, 3 Nov 2024 10:35:17 -0800 Subject: [PATCH 02/15] waiting on test run for sample outputs --- docs/index.rst | 82 ++++++++++++++++++++---------------------- docs/rqc_workflow.png | Bin 38172 -> 0 bytes rqcfilter.wdl | 13 +++---- 3 files changed, 45 insertions(+), 50 deletions(-) delete mode 100644 docs/rqc_workflow.png diff --git a/docs/index.rst b/docs/index.rst index b055f3c..7a275cf 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -1,7 +1,7 @@ -Reads QC Workflow (v1.0.2) +MetaT Reads QC Workflow (v0.0.7) ============================= -.. image:: rqc_workflow.png +.. image:: mt_rqc_workflow2024.png :align: center :scale: 50% @@ -17,7 +17,7 @@ The following parameters are used for "rqcfilter2" in this workflow: - clumpify=true : Run clumpify; all deduplication flags require this. - extend=false : Extend reads during merging to allow insert size estimation of non-overlapping reads. - jni=true : Enable C code for higher speed and identical results. - - usejni=false : Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code; details are in /jni/README.txt. + - usejni=false : Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code. - khist=true : Generate a kmer-frequency histogram of the output data. - maq=10 : Reads with average quality (before trimming) below this will be discarded. - maxns=1 : Reads with more Ns than this will be discarded. @@ -46,7 +46,7 @@ Workflow Availability The workflow from GitHub uses all the listed docker images to run all third-party tools. The workflow is available in GitHub: https://github.com/microbiomedata/metaT_ReadsQC; the corresponding -Docker image is available in DockerHub: https://hub.docker.com/r/microbiomedata/bbtools. +Docker images are available in DockerHub: https://hub.docker.com/r/microbiomedata/bbtools, https://hub.docker.com/r/microbiomedata/workflowmeta. Requirements for Execution -------------------------- @@ -87,7 +87,8 @@ The following commands will download the database:: Sample dataset(s) ----------------- -- Bulk soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_122 (`SRR8552838 `_)` with `metadata available in the NMDC Data Portal `_ +- Metatranscriptome of soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_RNA_119 (`SRR11678315 `_) with `metadata available in the NMDC Data Portal `_. The zipped fastq file is available `here `_ + Inputs @@ -95,14 +96,12 @@ Inputs A JSON file containing the following information: -1. the path to the database -2. the path to the interleaved fastq file (input data) -3. the path to the output directory -4. input_interleaved (boolean) -5. forwards reads fastq file (when input_interleaved is false) -6. reverse reads fastq file (when input_interleaved is false) -7. (optional) parameters for memory -8. (optional) number of threads requested +1. the path to the database directory +2. the path to the fastq file(s) ([R1, R2] if not interleaved) +3. input_interleaved (boolean) +4. output file prefix +5. (optional) parameters for memory +6. (optional) number of threads requested An example input JSON file is shown below: @@ -110,59 +109,55 @@ An example input JSON file is shown below: .. code-block:: JSON { - "metaTReadsQC.input_files": ["/global/cfs/cdirs/m3408/test_data/metatranscriptome/antisense/52400.2.318150.GATACTGG-CCAGTATC.fastq.gz"], - "metaTReadsQC.proj":"nmdc:xxxxxxx", + "metaTReadsQC.input_files": ["https://portal.nersc.gov/project/m3408//test_data/metaT/SRR11678315.fastq.gz"], + "metaTReadsQC.proj":"SRR11678315-int-0.1", "metaTReadsQC.rqc_mem": 180, "metaTReadsQC.rqc_thr": 64, "metaTReadsQC.database": "/refdata/" } -.. note:: - - In an HPC environment, parallel processing allows for processing multiple samples. The "jgi_rqcfilter.input_files" parameter is an array data structure. It can be used for multiple samples as input separated by a comma (,). - Ex: "jgi_rqcfilter.input_files":[“first-int.fastq”,”second-int.fastq”] Output ------ -A directory named with the prefix of the FASTQ input file will be created and multiple output files are generated; the main QC FASTQ output is named prefix.anqdpht.fastq.gz. Using the dataset above as an example, the main output would be named SRR7877884-int-0.1.anqdpht.fastq.gz. Other files include statistics on the quality of the data; what was trimmed, detected, and filtered in the data; a status log, and a shell script documenting the steps implemented so the workflow can be reproduced. +In the workflow execution directories, there will be a folder called `filtered` containing all the below listed output files. The bolded outputs below will be copied over to the primary output folder for the full workflow, these are what are shown through the NMDC-EDGE website. The `rqcfilter2.sh` output is named `raw.anqdpht.fastq.gz`. Using the dataset above as an example, the main output would be renamed `SRR11678315-int-0.1.filtered.fastq.gz`. Other files include statistics on the quality of the data; what was trimmed, detected, and filtered in the data; a status log, and a shell script documenting the steps implemented so the workflow can be reproduced. An example output JSON file (filterStats.json) is shown below: .. code-block:: JSON - { - "inputReads": 331126, - "kfilteredBases": 138732, - "qfilteredReads": 0, - "ktrimmedReads": 478, - "outputBases": 1680724, - "ktrimmedBases": 25248, - "kfilteredReads": 926, - "qtrimmedBases": 0, - "outputReads": 11212, - "gcPolymerRatio": 0.182857, - "inputBases": 50000026, - "qtrimmedReads": 0, - "qfilteredBases": 0 - } - - -Below is an example of all the output directory files with descriptions to the right. + { + "inputReads": 331126, + "kfilteredBases": 138732, + "qfilteredReads": 0, + "ktrimmedReads": 478, + "outputBases": 1680724, + "ktrimmedBases": 25248, + "kfilteredReads": 926, + "qtrimmedBases": 0, + "outputReads": 11212, + "gcPolymerRatio": 0.182857, + "inputBases": 50000026, + "qtrimmedReads": 0, + "qfilteredBases": 0 + } + + +Below is an example of all the `filtered` output directory files from `rqcfilter2.sh` with descriptions to the right. The **bolded** files are selected for output through NMDC-EDGE. ==================================== ============================================================================ FileName Description ==================================== ============================================================================ -nmdc_xxxxxx.fastq.gz main output (clean data) +**raw.anqrpht.fastq.gz main output (clean data)** adaptersDetected.fa adapters detected and removed bhist.txt base composition histogram by position cardinality.txt estimation of the number of unique kmers commonMicrobes.txt detected common microbes file-list.txt output file list for rqcfilter2.sh -filterStats.txt summary statistics +**filterStats.txt summary statistics filterStats.json summary statistics in JSON format -filterStats2.txt more detailed summary statistics +filterStats2.txt more detailed summary statistics** gchist.txt GC content histogram human.fq.gz detected human sequence reads ihist_merge.txt insert size histogram @@ -191,7 +186,7 @@ synth2.fq.gz detected synthetic molecule (short contami Version History --------------- -- 1.0.2 (release date **04/09/2021**; previous versions: 1.0.1) +- 0.0.7 (release date **08/23/2024**; previous versions: 0.0.6) Point of contact @@ -199,5 +194,4 @@ Point of contact - Original author: Brian Bushnell -- Package maintainer: Chienchi Lo - +- Package maintainers: Chienchi Lo diff --git a/docs/rqc_workflow.png b/docs/rqc_workflow.png deleted file mode 100644 index 44dbe29d52c0ce0d8c525cfddec533b48f04a6b6..0000000000000000000000000000000000000000 GIT binary patch literal 0 HcmV?d00001 literal 38172 zcmZU*by$_p^FDqk>5!5RNhPHl1eES>kd*H3R78<(kW{+6Q9-)9K|nw_)S=_M$JhJw z`{(C%ad~(=&+hE*%suza?D$?)Sq2+}3Jt&7yqYLARZR{>kfujOz^8dUB#EUU zi^kk3F3CJ#F_M<%HewH*H8~ja8WD-?O7rO$J@p?I_Sj1V5`jM=<6Wx=|2_q;NlE_q z$vY+-@!z8(5_FV*AJ$&r!75~xZPrl6rz)O{+31UQz5~>{6&l?o-YUXpd-J)En2=y)hWG+x|1d3F|hhVz{=`y z{6&cpRe=K8-m1;Np=tNHrx{7Kl~wFoCOUKHn0^yrfH9R5d0tQO2IBma6)zA+PM`Jd 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prefix=sub(proj, ":", "_") - Boolean gcloud_env=false Array[File?] input_files String database="/refdata/" + String proj Int rqc_mem = 180 Int rqc_thr = 64 - Int interleave_mem = 10 + Int interleave_mem = 10 + Boolean gcloud_env=false + String prefix=sub(proj, ":", "_") + String bbtools_container="microbiomedata/bbtools@sha256:df16e8343661effe8060478d14a9b4620cf484efba0d4818a467276d4bb07a6d" + String workflow_container = "microbiomedata/workflowmeta:1.1.1" + } if (length(input_files) == 1) { From 804060166dd1ceb5cc82e77a42d372cc877fcd7c Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Sun, 3 Nov 2024 11:02:11 -0800 Subject: [PATCH 03/15] add rrna file to output list, update figure --- docs/index.rst | 3 ++- docs/mt_rqc_workflow2024.png | Bin 0 -> 605237 bytes 2 files changed, 2 insertions(+), 1 deletion(-) create mode 100644 docs/mt_rqc_workflow2024.png diff --git a/docs/index.rst b/docs/index.rst index 7a275cf..5f423b8 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -149,7 +149,8 @@ Below is an example of all the `filtered` output directory files from `rqcfilter ==================================== ============================================================================ FileName Description ==================================== ============================================================================ -**raw.anqrpht.fastq.gz main output (clean data)** +**raw.anqrpht.fastq.gz main output (clean data) +rRNA.fastq.gz filtered ribosomal reads** adaptersDetected.fa adapters detected and removed bhist.txt base composition histogram by position cardinality.txt estimation of the number of unique kmers diff --git a/docs/mt_rqc_workflow2024.png b/docs/mt_rqc_workflow2024.png new file mode 100644 index 0000000000000000000000000000000000000000..b6dd587623d908e045d52e64aefe5560fd18adad GIT binary patch literal 605237 zcmeFacUV++_CJ0VBW6{IAG0=^i6~Z3QF=2LLabvjieV*T+%k%86i}Swsl-GHk*E#os*H5S_uKd@A 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output json and sample data --- docs/index.rst | 33 +++++++++++++++++---------------- 1 file changed, 17 insertions(+), 16 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 5f423b8..4fe9e90 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -9,7 +9,7 @@ MetaT Reads QC Workflow (v0.0.7) Workflow Overview ----------------- -This workflow utilizes the program "rqcfilter2" from BBTools to perform quality control on raw Illumina reads. The workflow performs quality trimming, artifact removal, linker trimming, adapter trimming, and spike-in removal (using BBDuk), and performs human/cat/dog/mouse/microbe removal (using BBMap). +This workflow utilizes the program "rqcfilter2" from BBTools to perform quality control on raw Illumina reads. The workflow performs quality trimming, artifact removal, linker trimming, adapter trimming, and spike-in removal (using BBDuk), and performs human/cat/dog/mouse/microbe removal (using BBMap). It is a replicate of the `QA protocol `_ implemented at JGI. The following parameters are used for "rqcfilter2" in this workflow: - barcodefilter=false : Disable improper barcodes filter @@ -77,8 +77,9 @@ Requisite database The RQCFilterData Database must be downloaded and installed. This is a 106 GB tar file which includes reference datasets of artifacts, adapters, contaminants, the phiX genome, and some host genomes. -The following commands will download the database:: +The following commands will download the database +.. code-block::bash mkdir refdata wget http://portal.nersc.gov/dna/microbial/assembly/bushnell/RQCFilterData.tar tar -xvf RQCFilterData.tar -C refdata @@ -87,7 +88,7 @@ The following commands will download the database:: Sample dataset(s) ----------------- -- Metatranscriptome of soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_RNA_119 (`SRR11678315 `_) with `metadata available in the NMDC Data Portal `_. The zipped fastq file is available `here `_ +- Metatranscriptome of soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_RNA_119 (`SRR11678315 `_) with `metadata available in the NMDC Data Portal `_. The zipped fastq file is available `here `_, and the sample outputs are available `here `_. @@ -128,19 +129,19 @@ An example output JSON file (filterStats.json) is shown below: .. code-block:: JSON { - "inputReads": 331126, - "kfilteredBases": 138732, - "qfilteredReads": 0, - "ktrimmedReads": 478, - "outputBases": 1680724, - "ktrimmedBases": 25248, - "kfilteredReads": 926, - "qtrimmedBases": 0, - "outputReads": 11212, - "gcPolymerRatio": 0.182857, - "inputBases": 50000026, - "qtrimmedReads": 0, - "qfilteredBases": 0 + "inputReads": 16809276, + "kfilteredBases": 4500, + "qfilteredReads": 3978, + "ktrimmedReads": 467761, + "outputBases": 1473400259, + "ktrimmedBases": 60463632, + "kfilteredReads": 15, + "qtrimmedBases": 2345, + "outputReads": 4974016, + "gcPolymerRatio": 112.898477, + "inputBases": 5076401352, + "qtrimmedReads": 292, + "qfilteredBases": 1185765 } From e8b4aa3867050a6cd37cbc311357dfb93eff7e37 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Thu, 14 Nov 2024 09:21:18 -0800 Subject: [PATCH 05/15] add svg --- docs/index.rst | 2 +- docs/mt_rqc_workflow2024.svg | 91 ++++++++++++++++++++++++++++++++++++ 2 files changed, 92 insertions(+), 1 deletion(-) create mode 100644 docs/mt_rqc_workflow2024.svg diff --git a/docs/index.rst b/docs/index.rst index 4fe9e90..f9b6035 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -1,7 +1,7 @@ MetaT Reads QC Workflow (v0.0.7) ============================= -.. image:: mt_rqc_workflow2024.png +.. image:: mt_rqc_workflow2024.svg :align: center :scale: 50% diff --git a/docs/mt_rqc_workflow2024.svg b/docs/mt_rqc_workflow2024.svg new file mode 100644 index 0000000..bda24e4 --- /dev/null +++ b/docs/mt_rqc_workflow2024.svg @@ -0,0 +1,91 @@ + + + + + + + + + + + + + + + + + + + + + rqcfilter2 + + + + RawReads + Pairednon-rRNAReads + + + + + + + + + + + + + \ No newline at end of file From 57b4360f378af0a2375a982c256392424a7f1129 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Thu, 14 Nov 2024 09:25:10 -0800 Subject: [PATCH 06/15] update readme with sample file link --- README.md | 15 +++++++++++---- 1 file changed, 11 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index e775c71..3bfcfb5 100644 --- a/README.md +++ b/README.md @@ -31,13 +31,20 @@ Description of the files: ## Input files -1. fastq (illumina paired-end interleaved fastq), -3. project name +1. the path to the database directory +2. the path to the fastq file(s) ([R1, R2] if not interleaved) +3. input_interleaved (boolean) +4. output file prefix +5. (optional) parameters for memory +6. (optional) number of threads requested ``` { - "metaTReadsQC.input_files": ["/global/cfs/cdirs/m3408/ficus/example/12889.1.295318.GTGCTTAC-GTAAGCAC.fastq.gz"], - "metaTReadsQC.proj":"nmdc:xxxxxxx" + "metaTReadsQC.input_files": ["https://portal.nersc.gov/project/m3408//test_data/metaT/SRR11678315.fastq.gz"], + "metaTReadsQC.proj":"SRR11678315-int-0.1", + "metaTReadsQC.rqc_mem": 180, + "metaTReadsQC.rqc_thr": 64, + "metaTReadsQC.database": "/refdata/" } ``` From 2e04b2d8b179ca031196b2af98f0423e4e269907 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Thu, 14 Nov 2024 16:22:05 -0800 Subject: [PATCH 07/15] updated sample datasets --- docs/index.rst | 7 +++---- 1 file changed, 3 insertions(+), 4 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index f9b6035..3c41394 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -87,10 +87,9 @@ The following commands will download the database Sample dataset(s) ----------------- - -- Metatranscriptome of soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_RNA_119 (`SRR11678315 `_) with `metadata available in the NMDC Data Portal `_. The zipped fastq file is available `here `_, and the sample outputs are available `here `_. - - +- Processed Metatranscriptome of soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_RNA_119 (`SRR11678315 `_) with `metadata available in the NMDC Data Portal `_. + - The zipped raw fastq file is available `here `_ + - The qc'ed outputs are available `here `_ Inputs ------ From c452fffac92dbba52884c5e3a0757c4b8f235121 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:09:36 -0800 Subject: [PATCH 08/15] change bold to italics and changed table format --- docs/index.rst | 119 +++++++++++++++++++++++++++++++------------------ 1 file changed, 76 insertions(+), 43 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 3c41394..586c3df 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -33,7 +33,7 @@ The following parameters are used for "rqcfilter2" in this workflow: - removemicrobes=true : Remove common contaminant microbial reads via mapping, and place them in a separate file. - removemouse=true : Remove mouse reads via mapping. - removeribo=true : Remove ribosomal reads via kmer-matching, and place them in a separate file. - - **rna=true** : Parameter for RNA-seq analysis. + - *rna=true* : Parameter for RNA-seq analysis. - sketch=true : Run SendSketch on 2M read pairs. - trimfragadapter=true: Trim all known Illumina adapter sequences, including TruSeq and Nextera. - trimq=0 : Trim quality threshold. @@ -51,10 +51,10 @@ Docker images are available in DockerHub: https://hub.docker.com/r/microbiomedat Requirements for Execution -------------------------- -(recommendations are in **bold**) +(recommendations are in *italics*) -- WDL-capable Workflow Execution Tool (**Cromwell**) -- Container Runtime that can load Docker images (**Docker v2.1.0.3 or higher**) +- WDL-capable Workflow Execution Tool (*Cromwell*) +- Container Runtime that can load Docker images (*Docker v2.1.0.3 or higher*) Hardware Requirements --------------------- @@ -144,50 +144,83 @@ An example output JSON file (filterStats.json) is shown below: } -Below is an example of all the `filtered` output directory files from `rqcfilter2.sh` with descriptions to the right. The **bolded** files are selected for output through NMDC-EDGE. - -==================================== ============================================================================ -FileName Description -==================================== ============================================================================ -**raw.anqrpht.fastq.gz main output (clean data) -rRNA.fastq.gz filtered ribosomal reads** -adaptersDetected.fa adapters detected and removed -bhist.txt base composition histogram by position -cardinality.txt estimation of the number of unique kmers -commonMicrobes.txt detected common microbes -file-list.txt output file list for rqcfilter2.sh -**filterStats.txt summary statistics -filterStats.json summary statistics in JSON format -filterStats2.txt more detailed summary statistics** -gchist.txt GC content histogram -human.fq.gz detected human sequence reads -ihist_merge.txt insert size histogram -khist.txt kmer-frequency histogram -kmerStats1.txt synthetic molecule (phix, linker, lamda, pJET) filter run log -kmerStats2.txt synthetic molecule (short contamination) filter run log -ktrim_kmerStats1.txt detected adapters filter run log -ktrim_scaffoldStats1.txt detected adapters filter statistics -microbes.fq.gz detected common microbes sequence reads -microbesUsed.txt common microbes list for detection -peaks.txt number of unique kmers in each peak on the histogram -phist.txt polymer length histogram -refStats.txt human reads filter statistics -reproduce.sh the shell script to reproduce the run -scaffoldStats1.txt detected synthetic molecule (phix, linker, lamda, pJET) statistics -scaffoldStats2.txt detected synthetic molecule (short contamination) statistics -scaffoldStatsSpikein.txt detected skipe-in kapa tag statistics -sketch.txt mash type sketch scanned result against nt, refseq, silva database sketches. -spikein.fq.gz detected skipe-in kapa tag sequence reads -status.log rqcfilter2.sh running log -synth1.fq.gz detected synthetic molecule (phix, linker, lamda, pJET) sequence reads -synth2.fq.gz detected synthetic molecule (short contamination) sequence reads -==================================== ============================================================================ +Below is an example of all the `filtered` output directory files from `rqcfilter2.sh` with descriptions to the right. The *italicized* files are selected for output through NMDC-EDGE. + +.. list-table:: + :header-rows: 1 + + * - Directory/File Name + - Description + * - *raw.anqrpht.fastq.gz* + - main output (clean data) + * - rRNA.fastq.gz + - filtered ribosomal reads + * - adaptersDetected.fa + - adapters detected and removed + * - bhist.txt + - base composition histogram by position + * - cardinality.txt + - estimation of the number of unique kmers + * - commonMicrobes.txt + - detected common microbes + * - file-list.txt + - output file list for rqcfilter2.sh + * - *filterStats.txt* + - *summary statistics* + * - *filterStats.json* + - *summary statistics in JSON format* + * - *filterStats2.txt* + - *more detailed summary statistics* + * - gchist.txt + - GC content histogram + * - human.fq.gz + - detected human sequence reads + * - ihist_merge.txt + - insert size histogram + * - khist.txt + - kmer-frequency histogram + * - kmerStats1.txt + - synthetic molecule (phix, linker, lamda, pJET) filter run log + * - kmerStats2.txt + - synthetic molecule (short contamination) filter run log + * - ktrim_kmerStats1.txt + - detected adapters filter run log + * - ktrim_scaffoldStats1.txt + - detected adapters filter statistics + * - microbes.fq.gz + - detected common microbes sequence reads + * - microbesUsed.txt + - common microbes list for detection + * - peaks.txt + - number of unique kmers in each peak on the histogram + * - phist.txt + - polymer length histogram + * - refStats.txt + - human reads filter statistics + * - reproduce.sh + - the shell script to reproduce the run + * - scaffoldStats1.txt + - detected synthetic molecule (phix, linker, lamda, pJET) statistics + * - scaffoldStats2.txt + - detected synthetic molecule (short contamination) statistics + * - scaffoldStatsSpikein.txt + - detected spike-in kapa tag statistics + * - sketch.txt + - mash type sketch scanned result against nt, refseq, silva database sketches + * - spikein.fq.gz + - detected spike-in kapa tag sequence reads + * - status.log + - rqcfilter2.sh running log + * - synth1.fq.gz + - detected synthetic molecule (phix, linker, lamda, pJET) sequence reads + * - synth2.fq.gz + - detected synthetic molecule (short contamination) sequence reads Version History --------------- -- 0.0.7 (release date **08/23/2024**; previous versions: 0.0.6) +- 0.0.7 (release date *08/23/2024*; previous versions: 0.0.6) Point of contact From 3733704806a34fd5d6970105aa21b4cc3b7e012b Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:17:34 -0800 Subject: [PATCH 09/15] converted parameter list to table and numbers to auto list --- docs/index.rst | 99 +++++++++++++++++++++++++++++++++----------------- 1 file changed, 66 insertions(+), 33 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 586c3df..bc17033 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -12,33 +12,66 @@ Workflow Overview This workflow utilizes the program "rqcfilter2" from BBTools to perform quality control on raw Illumina reads. The workflow performs quality trimming, artifact removal, linker trimming, adapter trimming, and spike-in removal (using BBDuk), and performs human/cat/dog/mouse/microbe removal (using BBMap). It is a replicate of the `QA protocol `_ implemented at JGI. The following parameters are used for "rqcfilter2" in this workflow: - - barcodefilter=false : Disable improper barcodes filter - - chastityfilter=false: Remove illumina reads failing chastity filter. - - clumpify=true : Run clumpify; all deduplication flags require this. - - extend=false : Extend reads during merging to allow insert size estimation of non-overlapping reads. - - jni=true : Enable C code for higher speed and identical results. - - usejni=false : Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code. - - khist=true : Generate a kmer-frequency histogram of the output data. - - maq=10 : Reads with average quality (before trimming) below this will be discarded. - - maxns=1 : Reads with more Ns than this will be discarded. - - minlen=51 : Reads shorter than this after trimming will be discarded. Pairs will be discarded only if both are shorter. - - mlf=0.33 : Reads shorter than this fraction of original length after trimming will be discarded. - - mtst=true : Spike-in bbduk removal mtst parameter - - phix=true : Remove reads containing phiX kmers. - - pigz=true : Use pigz for compression - - qtrim=r : Quality-trim from right ends before mapping. - - removecat=true : Remove cat reads via mapping. - - removedog=true : Remove dog reads via mapping. - - removehuman=true : Remove human reads via mapping. - - removemicrobes=true : Remove common contaminant microbial reads via mapping, and place them in a separate file. - - removemouse=true : Remove mouse reads via mapping. - - removeribo=true : Remove ribosomal reads via kmer-matching, and place them in a separate file. - - *rna=true* : Parameter for RNA-seq analysis. - - sketch=true : Run SendSketch on 2M read pairs. - - trimfragadapter=true: Trim all known Illumina adapter sequences, including TruSeq and Nextera. - - trimq=0 : Trim quality threshold. - - trimpolyg=5 : Trim reads that start or end with a G polymer at least this long. - - unpigz=t : Use pigz for decompression + +.. list-table:: + :header-rows: 1 + + * - `Parameter` + - Description + * - `barcodefilter=false` + - Disable improper barcodes filter + * - `chastityfilter=false` + - Remove illumina reads failing chastity filter + * - `clumpify=true` + - Run clumpify; all deduplication flags require this + * - `extend=false` + - Extend reads during merging to allow insert size estimation of non-overlapping reads + * - `jni=true` + - Enable C code for higher speed and identical results + * - `usejni=false` + - Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code + * - `khist=true` + - Generate a kmer-frequency histogram of the output data + * - `maq=10` + - Reads with average quality (before trimming) below this will be discarded + * - `maxns=1` + - Reads with more Ns than this will be discarded + * - `minlen=51` + - Reads shorter than this after trimming will be discarded. Pairs will be discarded only if both are shorter + * - `mlf=0.33` + - Reads shorter than this fraction of original length after trimming will be discarded + * - `mtst=true` + - Spike-in bbduk removal mtst parameter + * - `phix=true` + - Remove reads containing phiX kmers + * - `pigz=true` + - Use pigz for compression + * - `qtrim=r` + - Quality-trim from right ends before mapping + * - `removecat=true` + - Remove cat reads via mapping + * - `removedog=true` + - Remove dog reads via mapping + * - `removehuman=true` + - Remove human reads via mapping + * - `removemicrobes=true` + - Remove common contaminant microbial reads via mapping, and place them in a separate file + * - `removemouse=true` + - Remove mouse reads via mapping + * - `removeribo=true` + - Remove ribosomal reads via kmer-matching, and place them in a separate file + * - `*rna=true*` + - Parameter for RNA-seq analysis + * - `sketch=true` + - Run SendSketch on 2M read pairs + * - `trimfragadapter=true` + - Trim all known Illumina adapter sequences, including TruSeq and Nextera + * - `trimq=0` + - Trim quality threshold + * - `trimpolyg=5` + - Trim reads that start or end with a G polymer at least this long + * - `unpigz=t` + - Use pigz for decompression Workflow Availability @@ -96,12 +129,12 @@ Inputs A JSON file containing the following information: -1. the path to the database directory -2. the path to the fastq file(s) ([R1, R2] if not interleaved) -3. input_interleaved (boolean) -4. output file prefix -5. (optional) parameters for memory -6. (optional) number of threads requested +#. the path to the database directory +#. the path to the fastq file(s) ([R1, R2] if not interleaved) +#. input_interleaved (boolean) +#. output file prefix +#. (optional) parameters for memory +#. (optional) number of threads requested An example input JSON file is shown below: From 9fcd003b2ff3dea0452d884f3448276953b99ff9 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:25:41 -0800 Subject: [PATCH 10/15] testing inline code block --- docs/index.rst | 58 +++++++++++++++++++++++++------------------------- 1 file changed, 29 insertions(+), 29 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index bc17033..faf98a5 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -16,61 +16,61 @@ The following parameters are used for "rqcfilter2" in this workflow: .. list-table:: :header-rows: 1 - * - `Parameter` + * - Parameter - Description - * - `barcodefilter=false` + * - barcodefilter=false - Disable improper barcodes filter - * - `chastityfilter=false` + * - chastityfilter=false - Remove illumina reads failing chastity filter - * - `clumpify=true` + * - clumpify=true - Run clumpify; all deduplication flags require this - * - `extend=false` + * - extend=false - Extend reads during merging to allow insert size estimation of non-overlapping reads - * - `jni=true` + * - jni=true - Enable C code for higher speed and identical results - * - `usejni=false` + * - usejni=false - Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code - * - `khist=true` + * - khist=true - Generate a kmer-frequency histogram of the output data - * - `maq=10` + * - maq=10 - Reads with average quality (before trimming) below this will be discarded - * - `maxns=1` + * - maxns=1 - Reads with more Ns than this will be discarded - * - `minlen=51` + * - minlen=51 - Reads shorter than this after trimming will be discarded. Pairs will be discarded only if both are shorter - * - `mlf=0.33` + * - mlf=0.33 - Reads shorter than this fraction of original length after trimming will be discarded - * - `mtst=true` + * - mtst=true - Spike-in bbduk removal mtst parameter - * - `phix=true` + * - phix=true - Remove reads containing phiX kmers - * - `pigz=true` + * - pigz=true - Use pigz for compression - * - `qtrim=r` + * - qtrim=r - Quality-trim from right ends before mapping - * - `removecat=true` + * - removecat=true - Remove cat reads via mapping - * - `removedog=true` + * - removedog=true - Remove dog reads via mapping - * - `removehuman=true` + * - removehuman=true - Remove human reads via mapping - * - `removemicrobes=true` + * - removemicrobes=true - Remove common contaminant microbial reads via mapping, and place them in a separate file - * - `removemouse=true` + * - removemouse=true - Remove mouse reads via mapping - * - `removeribo=true` + * - removeribo=true - Remove ribosomal reads via kmer-matching, and place them in a separate file - * - `*rna=true*` + * - *rna=true* - Parameter for RNA-seq analysis - * - `sketch=true` + * - sketch=true - Run SendSketch on 2M read pairs - * - `trimfragadapter=true` + * - trimfragadapter=true - Trim all known Illumina adapter sequences, including TruSeq and Nextera - * - `trimq=0` + * - trimq=0 - Trim quality threshold - * - `trimpolyg=5` + * - trimpolyg=5 - Trim reads that start or end with a G polymer at least this long - * - `unpigz=t` + * - unpigz=t - Use pigz for decompression @@ -154,7 +154,7 @@ An example input JSON file is shown below: Output ------ -In the workflow execution directories, there will be a folder called `filtered` containing all the below listed output files. The bolded outputs below will be copied over to the primary output folder for the full workflow, these are what are shown through the NMDC-EDGE website. The `rqcfilter2.sh` output is named `raw.anqdpht.fastq.gz`. Using the dataset above as an example, the main output would be renamed `SRR11678315-int-0.1.filtered.fastq.gz`. Other files include statistics on the quality of the data; what was trimmed, detected, and filtered in the data; a status log, and a shell script documenting the steps implemented so the workflow can be reproduced. +In the workflow execution directories, there will be a folder called :code:`filtered` containing all the below listed output files. The bolded outputs below will be copied over to the primary output folder for the full workflow, these are what are shown through the NMDC-EDGE website. The :code:`rqcfilter2.sh` output is named :code:`raw.anqdpht.fastq.gz`. Using the dataset above as an example, the main output would be renamed :code:`SRR11678315-int-0.1.filtered.fastq.gz`. Other files include statistics on the quality of the data; what was trimmed, detected, and filtered in the data; a status log, and a shell script documenting the steps implemented so the workflow can be reproduced. An example output JSON file (filterStats.json) is shown below: From 20c497d3336431c66d552bf95f889eef475a4fa2 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:27:15 -0800 Subject: [PATCH 11/15] added :code: for in-line code --- docs/index.rst | 58 ++++++++++++++++++++++++-------------------------- 1 file changed, 28 insertions(+), 30 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index faf98a5..e76f24f 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -16,61 +16,59 @@ The following parameters are used for "rqcfilter2" in this workflow: .. list-table:: :header-rows: 1 - * - Parameter - - Description - * - barcodefilter=false + * - :code:`barcodefilter=false` - Disable improper barcodes filter - * - chastityfilter=false + * - :code:`chastityfilter=false` - Remove illumina reads failing chastity filter - * - clumpify=true + * - :code:`clumpify=true` - Run clumpify; all deduplication flags require this - * - extend=false + * - :code:`extend=false` - Extend reads during merging to allow insert size estimation of non-overlapping reads - * - jni=true + * - :code:`jni=true` - Enable C code for higher speed and identical results - * - usejni=false + * - :code:`usejni=false` - Do alignments in C code, which is faster, if an edit distance is allowed. This will require compiling the C code - * - khist=true + * - :code:`khist=true` - Generate a kmer-frequency histogram of the output data - * - maq=10 + * - :code:`maq=10` - Reads with average quality (before trimming) below this will be discarded - * - maxns=1 + * - :code:`maxns=1` - Reads with more Ns than this will be discarded - * - minlen=51 + * - :code:`minlen=51` - Reads shorter than this after trimming will be discarded. Pairs will be discarded only if both are shorter - * - mlf=0.33 + * - :code:`mlf=0.33` - Reads shorter than this fraction of original length after trimming will be discarded - * - mtst=true + * - :code:`mtst=true` - Spike-in bbduk removal mtst parameter - * - phix=true + * - :code:`phix=true` - Remove reads containing phiX kmers - * - pigz=true + * - :code:`pigz=true` - Use pigz for compression - * - qtrim=r + * - :code:`qtrim=r` - Quality-trim from right ends before mapping - * - removecat=true + * - :code:`removecat=true` - Remove cat reads via mapping - * - removedog=true + * - :code:`removedog=true` - Remove dog reads via mapping - * - removehuman=true + * - :code:`removehuman=true` - Remove human reads via mapping - * - removemicrobes=true + * - :code:`removemicrobes=true` - Remove common contaminant microbial reads via mapping, and place them in a separate file - * - removemouse=true + * - :code:`removemouse=true` - Remove mouse reads via mapping - * - removeribo=true + * - :code:`removeribo=true` - Remove ribosomal reads via kmer-matching, and place them in a separate file - * - *rna=true* + * - :code:`*rna=true*` - Parameter for RNA-seq analysis - * - sketch=true + * - :code:`sketch=true` - Run SendSketch on 2M read pairs - * - trimfragadapter=true + * - :code:`trimfragadapter=true` - Trim all known Illumina adapter sequences, including TruSeq and Nextera - * - trimq=0 + * - :code:`trimq=0` - Trim quality threshold - * - trimpolyg=5 + * - :code:`trimpolyg=5` - Trim reads that start or end with a G polymer at least this long - * - unpigz=t + * - :code:`unpigz=t` - Use pigz for decompression @@ -177,7 +175,7 @@ An example output JSON file (filterStats.json) is shown below: } -Below is an example of all the `filtered` output directory files from `rqcfilter2.sh` with descriptions to the right. The *italicized* files are selected for output through NMDC-EDGE. +Below is an example of all the :code:`filtered` output directory files from :code:`rqcfilter2.sh` with descriptions to the right. The *italicized* files are selected for output through NMDC-EDGE. .. list-table:: :header-rows: 1 From 6d5ce7f3326e06771f68d52eeb34be37ebad6fa2 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:27:56 -0800 Subject: [PATCH 12/15] added missing table header --- docs/index.rst | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/docs/index.rst b/docs/index.rst index e76f24f..0c1c648 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -15,7 +15,8 @@ The following parameters are used for "rqcfilter2" in this workflow: .. list-table:: :header-rows: 1 - + * - :code:`Parameter` + - Description * - :code:`barcodefilter=false` - Disable improper barcodes filter * - :code:`chastityfilter=false` From f41b08daea536973e1c475629a425736f4bb05e0 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:28:46 -0800 Subject: [PATCH 13/15] missing line break --- docs/index.rst | 1 + 1 file changed, 1 insertion(+) diff --git a/docs/index.rst b/docs/index.rst index 0c1c648..58241d8 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -15,6 +15,7 @@ The following parameters are used for "rqcfilter2" in this workflow: .. list-table:: :header-rows: 1 + * - :code:`Parameter` - Description * - :code:`barcodefilter=false` From c5fc2aa8c94d41471ccc35c7b886db6bf96763a1 Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Fri, 15 Nov 2024 10:30:25 -0800 Subject: [PATCH 14/15] added detail to difference wtih readsqc --- docs/index.rst | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 58241d8..268b075 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -15,7 +15,7 @@ The following parameters are used for "rqcfilter2" in this workflow: .. list-table:: :header-rows: 1 - + * - :code:`Parameter` - Description * - :code:`barcodefilter=false` @@ -60,8 +60,8 @@ The following parameters are used for "rqcfilter2" in this workflow: - Remove mouse reads via mapping * - :code:`removeribo=true` - Remove ribosomal reads via kmer-matching, and place them in a separate file - * - :code:`*rna=true*` - - Parameter for RNA-seq analysis + * - :code:`rna=true` + - *Parameter for RNA-seq analysis* (this is main difference between `ReadsQC `_ and MetaT ReadsQC) * - :code:`sketch=true` - Run SendSketch on 2M read pairs * - :code:`trimfragadapter=true` From b5a78ebf96fc6f35c48778259bacd78641b4487b Mon Sep 17 00:00:00 2001 From: Kaitlyn Jiayi Li Date: Mon, 18 Nov 2024 23:02:46 -0800 Subject: [PATCH 15/15] fix bullets --- docs/index.rst | 11 +++++++---- 1 file changed, 7 insertions(+), 4 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 268b075..68cefee 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -78,8 +78,10 @@ Workflow Availability --------------------- The workflow from GitHub uses all the listed docker images to run all third-party tools. -The workflow is available in GitHub: https://github.com/microbiomedata/metaT_ReadsQC; the corresponding -Docker images are available in DockerHub: https://hub.docker.com/r/microbiomedata/bbtools, https://hub.docker.com/r/microbiomedata/workflowmeta. +The workflow is available in GitHub: https://github.com/microbiomedata/metaT_ReadsQC; the corresponding Docker images are available in DockerHub: + +- `microbiomedata/bbtools:38.96 `_ +- `microbiomedata/workflowmeta:1.1.1 `_ Requirements for Execution -------------------------- @@ -118,9 +120,10 @@ The following commands will download the database tar -xvf RQCFilterData.tar -C refdata rm RQCFilterData.tar -Sample dataset(s) ------------------ +Sample datasets +--------------- - Processed Metatranscriptome of soil microbial communities from the East River watershed near Crested Butte, Colorado, United States - ER_RNA_119 (`SRR11678315 `_) with `metadata available in the NMDC Data Portal `_. + - The zipped raw fastq file is available `here `_ - The qc'ed outputs are available `here `_

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