Samtool error

[hsmart]

Hi All,
I am getting following error with SAMtools while converting .sam file to .bam file.
samtools view -bt hg19.fa Input.sam -o Output.bam
Parse error at line 103686571: sequence and quality are inconsistent
Is there any easy to fix this issue (i.e., remove the sequence lines that are not consistent with quality values)?
Thanks for your help,
Hemang

[Heisman]

Probably worthwhile to figure out why you're getting that problem in the first place. Can you copy and paste the read(s) that are causing the error? What sequencing platform are you using?

[hsmart]

Thanks for looking into it. We are using Illumina platform to generate data. I have used several packages (i.e., BWA, asSeq in R and samtools) so basically some of the reads were trimmed. Is there any easy to fix this issue (i.e., remove the sequence lines that are not consistent with quality values)? Here is the read that is causing the error:

HWI-ST165_0195:6:68:21171:200547#0 77 * 0 0 * * 0 0 CACCTATTCATACTCGTGCTCTGGCTCGGCAATCACCTGTAGCAGCAAAAACAAAGAGGGTGAGGCCTTCATT HHHHHHHHHHHHHHHHHHFDDDGFEFDHFHHED>HFD<C>7A>?A@EEAE2+9:1+&(%68<<@=CDEEE?AD RG:Z:111_N_R

Thanks for your help,

Hemang

[Heisman]

I think it might just be your command as I took that read and aligned it/converted it to a bam file without problems.

Maybe try the following if you also want to sort the file:

samtools view -uS Input.sam | samtools sort - Input_sorted

Otherwise try:

samtools view -bS Input.sam -o Output.bam

[hsmart]

Thank you so much for your help. It worked perfectly fine.