tophat strand info?

[frankyue50]

can anyone help me identify if the strand info is in the accepted.sam file (tophat output)
SOLEXA1_0001:1:60:7595:15462#0 16 chr1 3000555 255 36M * 0 0 AATTTGTTTTGTCCTTCAGCGGATTAAAATATTCAG CBBBCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC NM:i:0

or does anyone know what is column 2 and 4 means. I checked sam manual but still confused. Thanks.

[john_mu]

Hi,

The strand info will only be on the junction reads. It is impossible to determine the strand of non-junction hits without using a stranded protocol in the experiment.

column 2 is the SAM flag. You can google "SAM flag" to find out.

column 4 is the chromosome start location of the hit.

[smueller]

Hey,

I just came accross the same problem and found that you can read strand info from the
2nd colunmn. This is actually a flag that has to be deciphered according to the SAM format and can be done for instance here:

http://picard.sourceforge.net/explain-flags.html

In your case, 16 means that this read is on the reverse strand.
Column 4 is the start postition of this read on the respective chromosome.
Note, since tophat produce BAM formats as default for a while, you can still
access the flag information with:
samtools view accepted_hits.bam
which produces SAM format.

cheers
Sebastian