Entering edit mode
9.8 years ago
R
▴
10
Hi,
htseq-count only accept BAM/SAM. I downloaded a bed format RNA-seq data (strand-specific, single end). it has around 19 million tags. Bam file is not provided (only sra) and when I converted bed by bedToBam and ran htseq-count:
no_feature 17528277
ambiguous 1977
too_low_aQual 0
not_aligned 0
alignment_not_unique 0
a lots of reads discarded!!!
bed format:
chr1 100002008 100002043 0 0 -
chr1 100002027 100002062 0 0 -
bedToBam
0 16 chr1 100002009 255 35M * 0 0 * *
0 16 chr1 100002028 255 35M * 0 0 * *
I really appreciate if somebody could please help me
What does 0 0 means between end and strand. Was it introduced by error when typing ? Can you paste a few lines (excluding header) may be 2-3 from the BAM file that was generated by bedtoBam.
I added few lines from bed and bam
Try
-s reverse
and see if that improves things.I used it when I ran the command
Then try
-s yes
. It's likely that either the strandedness setting is incorrect or the annotation and reference against things were mapped don't match. It's usually easiest to just look at things in IGV.