Changes between Version 28 and Version 29 of SOPs/atac_Seq


Ignore:
Timestamp:
10/27/20 14:45:59 (4 years ago)
Author:
byuan
Comment:

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  • SOPs/atac_Seq

    v28 v29  
    7979
    8080The [https://www.encodeproject.org/atac-seq/#standards ENCODE project] has some recommended TSS enrichment scores, depending on the genome.
     81
     82In 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
     84First, run ataqv on each bam file to generate JSON files.
     85Here 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
     100ataqv --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
     105Run 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.
     110mkarv 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"
     113mkarv all_samples sample1.ataqv.json.gz sample2.ataqv.json.gz
     114
     115}}}
     116
    81117
    82118=== [=#call_peaks Call peaks] ===