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training:dudijon:galaxy [2021/12/14 18:26] slegras [7.2 Rename the workflow DNA-seq data analysis] |
training:dudijon:galaxy [2022/12/09 15:37] (current) slegras |
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^ Instructor ^ Stephanie Le Gras ^ | ^ Instructor ^ Stephanie Le Gras ^ | ||
^ Duration | 3.5 hours | | ^ Duration | 3.5 hours | | ||
- | ^ Content | {{:training:dudijon:introgalaxy_2020_compressed.pdf|Description of the key features of Galaxy (Lecture)}} | | + | ^ Content | {{:training:dudijon:introgalaxy_2021_compressed.pdf|Description of the key features of Galaxy (Lecture)}} | |
^ ::: | Practical session on basic features of Galaxy (Hands-on) | | ^ ::: | Practical session on basic features of Galaxy (Hands-on) | | ||
^ Prerequisites | None | | ^ Prerequisites | None | | ||
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- | ==== - Import data into Galaxy ==== | + | ==== - Import files from your computer to Galaxy ==== |
- | === - Import files from your computer to Galaxy === | + | |
- | - Download the file “**sample.bed.gz**” following this [[https://seafile.igbmc.fr/d/345d7581237d4295bf2c/|link]] and upload it to Galaxy. | + | - Download the file “**sample.bed.gz**” following this [[https://seafile.igbmc.fr/d/1adaad8f80394182a784/|link]] and upload it to Galaxy. |
* The genome is: Mouse (mm9) | * The genome is: Mouse (mm9) | ||
* The format is: bed | * The format is: bed | ||
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==== - Running a tool ==== | ==== - Running a tool ==== | ||
- | - Download the two files **CRN-107_11-R1.fastq.gz** and **CRN-107_11-R2.fastq.gz** following this [[https://seafile.igbmc.fr/d/345d7581237d4295bf2c/|link]]. | + | - Download the two files **CRN-107_11-R1.fastq.gz** and **CRN-107_11-R2.fastq.gz** following this [[https://seafile.igbmc.fr/d/1adaad8f80394182a784/|link]]. |
- Import them to your history called “DNA-seq data analysis” | - Import them to your history called “DNA-seq data analysis” | ||
* The genome is: Human (hg19) | * The genome is: Human (hg19) | ||
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* CRN-107_11-R1.fastq | * CRN-107_11-R1.fastq | ||
* CRN-107_11-R2.fastq | * CRN-107_11-R2.fastq | ||
- | * CaptureDesign_chr4.bed (download it from [[https://seafile.igbmc.fr/d/345d7581237d4295bf2c/|here]]) | + | * CaptureDesign_chr4.bed (download it from [[https://seafile.igbmc.fr/d/1adaad8f80394182a784/|here]]) |
Import missing files from the data library "**DNA-seq test datasets**" | Import missing files from the data library "**DNA-seq test datasets**" | ||
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{{:training:dudijon:05-editorrunworklow.png?|}} | {{:training:dudijon:05-editorrunworklow.png?|}} | ||
- | Now your can edit or run the workflow: | ||
- | |||
- | {{:training:dudijon:06-editworklow.png?|}} | ||
++++ | ++++ | ||
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- __Samtools flagstat__ to compute mapping statistics (after BWA mem) | - __Samtools flagstat__ to compute mapping statistics (after BWA mem) | ||
- | - __Filter__ to select aligned reads with a mapping quality >= 20 (after MarkDuplicates) | + | - __Filter SAM or BAM, output SAM or BAM__ to select aligned reads with a mapping quality >= 20 (after MarkDuplicates) |
- __Samtools flagstat__ to compute mapping statistics after removing reads with low mapping qualities (after Filter) | - __Samtools flagstat__ to compute mapping statistics after removing reads with low mapping qualities (after Filter) | ||
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- __Flagstat__ tabulate descriptive stats for BAM dataset | - __Flagstat__ tabulate descriptive stats for BAM dataset | ||
- BAM File to Convert: **output of BWA mem** | - BAM File to Convert: **output of BWA mem** | ||
- | - __Filter__ BAM datasets on a variety of attributes | + | - __Filter SAM or BAM, output SAM or BAM__ files on FLAG MAPQ RG LN or by region |
- | - BAM dataset(s) to filter: **output of Picard MarkDuplicates** | + | - SAM or BAM file to filter: **output of Picard MarkDuplicates** |
- | - Select BAM property to filter on: mapQuality | + | - Minimum MAPQ quality score: **20** |
- | - Filter on read mapping quality (phred scale): **>=20** (this exact expression, including ">="!) | + | |
- __Flagstat__ tabulate descriptive stats for BAM dataset | - __Flagstat__ tabulate descriptive stats for BAM dataset | ||
- BAM File to Convert: **output of Filter** | - BAM File to Convert: **output of Filter** | ||
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++++ Answer | | ++++ Answer | | ||
- | 561598 - 531417 = 30181 | + | 561598 - 530355 = 31243 |
++++ | ++++ | ||