Filtering uniquely mapping reads from bam files generated from STAR
0
0
Entering edit mode
3.4 years ago
Kai_Qi ▴ 130

I am learning to use MACS2 to call peaks for CLIP-Seq. I have bamfiles generated from STAR using the following command:

#get the input data file
    INPUT=${1}

    module load STAR
    STAR --genomeDir /scratch/midway2/caiqi/GRCh37_star_index_150bp \
    --runThreadN 16 \
    --readFilesIn ${INPUT}_1.fastq ${INPUT}_2.fastq \
    --outFileNamePrefix GRCh37${INPUT} \
    --twopassMode Basic \
    --sjdbOverhang 150 \
    --outSAMtype BAM SortedByCoordinate \
    --outFilterMultimapNmax 20 \
    --outFilterMismatchNmax 999 \
    --outFilterMismatchNoverLmax 0.06 \
     --alignIntronMin 70 \
     --alignIntronMax 500000 \
     --alignMatesGapMax 500000 \
     --alignSJoverhangMin 8 \
     --alignSJDBoverhangMin 1 \
     --outSAMstrandField intronMotif \
     --outFilterType BySJout
    (END)

Now I would like to filter the BAM files using

sambamba as advised:

$ sambamba view -h -t 2 -f bam \
-F "[XS] == null and not unmapped  and not duplicate" \
H1hesc_Input_Rep1_chr12_aln_sorted.bam > H1hesc_Input_Rep1_chr12_aln.bam

The parameter following -F is [XS] is not appropriate for me because ‘XS’ is a tag generated by Bowtie2 that gives an alignment score for the second-best alignment, and it is only present if the read is aligned and more than one alignment was found for the read.

How can modify the argument after -F to make it suitable for the bam files from STAR?

Thanks,

rna-seq RNA-Seq ChIP-Seq next-gen • 1.1k views
ADD COMMENT

Login before adding your answer.

Traffic: 1849 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6