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Checking whether the reads aligned to specific gene have the specific exon in BAM file

Hi guys. So I did a cellranger count on my sample with my own new gene in the gtf annotation file.

And from the bam file I found some of the reads aligned to the gene according to its GX tag.

This gene however must have the specific exon sequence expressed, so I want to double check whether these aligned reads do have this specific exon sequence.

Right now I managed to use samtools to select the reads with the GX:gene tag from the bam file. How can I check whether these reads have that specific exonic sequence?

Thanks a lot! Appreciate any help.

rna-seq sequencing alignment samtools

By loading that file into IGV ?

feels like there are far too many reads to check the exon in igv

1 answer

One option is to get the subset of bam file mapping to that region:

samtools view -b in.bam "chr:1000-100000" > in.subset.bam and view in IGV.

Other option is to get exonic counts using featureCounts.

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