| 81 | |
| 82 | In addition to do varies quality controls, [[ https://www.sciencedirect.com/science/article/pii/S240547122030079X | ataqv ]] summarizes QC results into an interactive html page, which also allows you to view multiple samples together. |
| 83 | |
| 84 | First, run ataqv on each bam file to generate JSON files. |
| 85 | Here is a sample command for a bulk ATAC_seq sample: |
| 86 | |
| 87 | |
| 88 | {{{ |
| 89 | |
| 90 | # --peak-file: peak file in bed format |
| 91 | # --tss-file: tss in bed format |
| 92 | # --excluded-region-file A BED file containing excluded regions, in this case, we exclude the regions in the ENCODE blast list |
| 93 | # --metrics-file: output in json format |
| 94 | # --ignore-read-groups: Even if read groups are present in the BAM file, ignore them and combine metrics for all reads under a single sample and library named with the --name option. This also implies that a single peak file will be used for all reads. |
| 95 | # The duplicated reads need to be labeled by Picard MarkDuplicates ( REMOVE_DUPLICATES=FALSE) before running the ataqv |
| 96 | # ataqc supports human/mouse/rat/fly/worm/yeast currently. If your genome is not listed, you can run it with --autosomal-reference-file and --mitochondrial-reference-name |
| 97 | # --autosomal-reference-file: a file containing autosomal reference names, one per line |
| 98 | # --mitochondrial-reference-name: name for the mitochondrial DNA |
| 99 | |
| 100 | ataqv --peak-file sample1_peak.bed --name sample1 --metrics-file sample1.ataqv.json.gz --excluded-region-file /nfs/genomes/human_hg38_dec13/anno/hg38-blacklist.v2.bed.gz --tss-file hg38.tss.refseq.bed.gz --ignore-read-groups human sample1.bam > sample1.ataqv.out” |
| 101 | |
| 102 | }}} |
| 103 | |
| 104 | |
| 105 | Run mkarv on the JSON files to generate the interactive web viewer. By default, SRR891268 will be used as the reference sample in the viewer. You can specify a different reference when you built your viewer instance, refer to the information on how to configure the reference with mkarv -h. |
| 106 | |
| 107 | {{{ |
| 108 | |
| 109 | # This will create a folder named as sample1, whose index.html contains the interactive plots. |
| 110 | mkarv sample1 sample1.ataqv.json.gz |
| 111 | |
| 112 | # I you have multiple samples, you can combine them into a single report. In this case, both sample1 and sample2 will be in the same plots inside folder called "all_samples" |
| 113 | mkarv all_samples sample1.ataqv.json.gz sample2.ataqv.json.gz |
| 114 | |
| 115 | }}} |
| 116 | |