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Merge pull request #121 from bnovak32/patch-4
MultiQC correction, typo fixes
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Metagenomics/Illumina/Pipeline_GL-DPPD-7107_Versions/GL-DPPD-7107-A.md

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@@ -68,17 +68,17 @@ Lauren Sanders (OSDR Project Scientist)
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|FastQC| 0.12.1 |[https://www.bioinformatics.babraham.ac.uk/projects/fastqc/](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)|
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|MultiQC| 1.19 |[https://multiqc.info/](https://multiqc.info/)|
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|bbduk| 38.86 |[https://jgi.doe.gov/data-and-tools/software-tools/bbtools/bb-tools-user-guide/](https://jgi.doe.gov/data-and-tools/software-tools/bbtools/bb-tools-user-guide/)|
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|megahit| 1.2.9 |[https://github.com/voutcn/megahit#megahit](https://github.com/voutcn/megahit#megahit)|
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|MEGAHIT| 1.2.9 |[https://github.com/voutcn/megahit#megahit](https://github.com/voutcn/megahit#megahit)|
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|bit| 1.8.53 |[https://github.com/AstrobioMike/bioinf_tools#bioinformatics-tools-bit](https://github.com/AstrobioMike/bioinf_tools#bioinformatics-tools-bit)|
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|bowtie2| 2.4.1 |[https://github.com/BenLangmead/bowtie2#overview](https://github.com/BenLangmead/bowtie2#overview)|
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|samtools| 1.20 |[https://github.com/samtools/samtools#samtools](https://github.com/samtools/samtools#samtools)|
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|prodigal| 2.6.3 |[https://github.com/hyattpd/Prodigal#prodigal](https://github.com/hyattpd/Prodigal#prodigal)|
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|Prodigal| 2.6.3 |[https://github.com/hyattpd/Prodigal#prodigal](https://github.com/hyattpd/Prodigal#prodigal)|
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|KOFamScan| 1.3.0 |[https://github.com/takaram/kofam_scan#kofamscan](https://github.com/takaram/kofam_scan#kofamscan)|
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|CAT| 5.2.3 |[https://github.com/dutilh/CAT#cat-and-bat](https://github.com/dutilh/CAT#cat-and-bat)|
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|MetaBAT| 2.15 |[https://bitbucket.org/berkeleylab/metabat/src/master/](https://bitbucket.org/berkeleylab/metabat/src/master/)|
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|checkm| 1.1.3 |[https://github.com/Ecogenomics/CheckM](https://github.com/Ecogenomics/CheckM)|
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|CheckM| 1.1.3 |[https://github.com/Ecogenomics/CheckM](https://github.com/Ecogenomics/CheckM)|
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|GTDB-Tk| 2.4.0 |[https://github.com/Ecogenomics/GTDBTk](https://github.com/Ecogenomics/GTDBTk)|
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|KEGGDecoder| 1.2.2 |[https://github.com/bjtully/BioData/tree/master/KEGGDecoder#kegg-decoder](https://github.com/bjtully/BioData/tree/master/KEGGDecoder#kegg-decoder)
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|KEGG-Decoder| 1.2.2 |[https://github.com/bjtully/BioData/tree/master/KEGGDecoder#kegg-decoder](https://github.com/bjtully/BioData/tree/master/KEGGDecoder#kegg-decoder)
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|HUMAnN| 3.9 |[https://github.com/biobakery/humann](https://github.com/biobakery/humann)|
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|MetaPhlAn| 4.1.0 |[https://github.com/biobakery/MetaPhlAn](https://github.com/biobakery/MetaPhlAn)|
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@@ -113,7 +113,7 @@ fastqc -o raw_fastqc_output *raw.fastq.gz
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#### 1a. Compile Raw Data QC
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```
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multiqc -o raw_multiqc_output -n raw_multiqc -z raw_fastqc_output/
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multiqc -o raw_multiqc_output -n raw_multiqc raw_fastqc_output/
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# this is how it's packaged with our workflow outputs
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zip -r raw_multiqc_GLmetagenomics_report.zip raw_multiqc_output
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```
@@ -122,7 +122,6 @@ zip -r raw_multiqc_GLmetagenomics_report.zip raw_multiqc_output
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* `-o` – the output directory to store results
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* `-n` – the filename prefix of results
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* `-z` – specifies to zip the output data directory
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* `raw_fastqc_output/` – the directory holding the output data from the fastqc run, provided as a positional argument
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**Input data:**
@@ -175,7 +174,7 @@ bbduk.sh in=sample-1-R1-raw.fastq.gz in2=sample-1-R2-raw.fastq.gz out1=sample-1_
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* `maxns` – sets the maximum number of Ns allowed in a read before it will be filtered out
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* `swift` – tells the program to look for and trim low-complexity adaptase reminants from the Swift1S kit
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* `swift` – tells the program to look for and trim low-complexity adaptase reminants from the Swift1S kit
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* `> bbduk.log 2>&1` – redirects the stderr and stdout to a log file for saving
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@@ -214,7 +213,7 @@ fastqc -o filtered_fastqc_output/ *filtered.fastq.gz
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#### 3a. Compile Filtered/Trimmed Data QC
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```
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multiqc -o filtered_multiqc_output -n filtered_multiqc -z filtered_fastqc_output/
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multiqc -o filtered_multiqc_output -n filtered_multiqc filtered_fastqc_output/
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# this is how it's packaged with our workflow outputs
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zip -r filtered_multiqc_GLmetagenomics_report.zip filtered_multiqc_output
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```
@@ -223,7 +222,6 @@ zip -r filtered_multiqc_GLmetagenomics_report.zip filtered_multiqc_output
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* `-o` – the output directory to store results
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* `-n` – the filename prefix of results
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* `-z` – specifies to zip the output data directory
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* `filtered_fastqc_output/` – the directory holding the output data from the fastqc run, provided as a positional argument
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**Input data:**
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### 4. Sample assembly
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```
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megahit -1 sample-1_R1_filtered.fastq.gz -2 sample-1_R2_filtered.fastq.gz \
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-o sample-1-assembly -t 10 --min-contig-length 500 > sample-1-assembly.log 2>&1
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-o sample-1-assembly -t NumberOfThreads --min-contig-length 500 > sample-1-assembly.log 2>&1
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```
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**Parameter Definitions:**
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#### 9b. Performing mapping, conversion to bam, and sorting
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```
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bowtie2 --threads 15 -x sample-1-assembly-bt-index -1 sample-1_R1_filtered.fastq.gz \
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-2 sample-1_R2_filtered.fastq.gz 2> sample-1-mapping-info.txt | samtools view -b | samtools sort -@ 15 > sample-1.bam
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bowtie2 --threads NumberOfThreads -x sample-1-assembly-bt-index -1 sample-1_R1_filtered.fastq.gz \
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-2 sample-1_R2_filtered.fastq.gz 2> sample-1-mapping-info.txt | samtools view -b | samtools sort -@ NumberOfThreads > sample-1.bam
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```
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**Parameter Definitions:**
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#### 9c. Indexing
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```
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samtools index -@ 15 sample-1.bam
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samtools index -@ NumberOfThreads sample-1.bam
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```
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**Parameter Definitions:**
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```
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jgi_summarize_bam_contig_depths --outputDepth sample-1-metabat-assembly-depth.tsv --percentIdentity 97 --minContigLength 1000 --minContigDepth 1.0 --referenceFasta sample-1-assembly.fasta sample-1.bam
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metabat2 --inFile sample-1-assembly.fasta --outFile sample-1 --abdFile sample-1-metabat-assembly-depth.tsv -t 4
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metabat2 --inFile sample-1-assembly.fasta --outFile sample-1 --abdFile sample-1-metabat-assembly-depth.tsv -t NumberOfThreads
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mkdir sample-1-bins
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mv sample-1*bin*.fasta sample-1-bins
@@ -993,7 +991,7 @@ metaphlan --install
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# forward and reverse reads need to be provided combined if paired-end (if not paired-end, single-end reads are provided to the --input argument next)
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cat sample-1_R1_filtered.fastq.gz sample-1_R2_filtered.fastq.gz > sample-1-combined.fastq.gz
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humann --input sample-1-combined.fastq.gz --output sample-1-humann3-out-dir --threads 15 \
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humann --input sample-1-combined.fastq.gz --output sample-1-humann3-out-dir --threads NumberOfThreads \
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--output-basename sample-1 --metaphlan-options "--unknown_estimation --add_viruses \
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--sample_id sample-1"
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```

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