Question: BLAST stranded SRA for plus or minus?
gravatar for Adria
7 weeks ago by
Adria10 wrote:

I'm trying to look into existing RNAseq datasets to see if my highly expressed gene of interest may have both plus and minus strands expressed. There are a pack of SRA datasets that are strand specific. I know that I can blast them (SRA nblast), but when I blast the known mRNA and the reverse complement I get the same pattern.

Any tips/tricks to get at this?

blast rna-seq sra • 148 views
ADD COMMENTlink modified 4 days ago by h.mon10k • written 7 weeks ago by Adria10
gravatar for h.mon
4 days ago by
h.mon10k wrote:

Map the SRA datasets with STAR to the appropriate annotated reference genome and using the --quantMode GeneCounts parameter. With this parameter, STAR will output gene counts for all strandedness options:

column 1: gene ID

column 2: counts for unstranded RNA-seq

column 3: counts for the 1st read strand aligned with RNA (htseq-count option -s yes)

column 4: counts for the 2nd read strand aligned with RNA (htseq-count option -s reverse)

Then you can:

  1. check the quality of strand-specificity with RSeQC infer-experiment-py
  2. look at your gene of interest counts
  3. view the bam mapping files with IGV
ADD COMMENTlink written 4 days ago by h.mon10k
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