Samtools extraction output is empty

Question

[spidcock01]

Sorry if similar questions have been answered already - I had a look and can't see anything that suits my problem exactly.

I'm trying to use Samtools to extract Fasta sequences from a database file using a list. (Sorry if my terminology is dreadful, I'm new to computational biology).

So I have my database file as a multi-FASTA file, and my list purely as a list of gene headers in a text file, e.g.

ND3005_accessory_2122_length_3432

etc. These do exactly match the headers in the database, with the exception of not having a > at the start of each sequence, but I've tried with these as well.

The only output that I get is a file with the headers in, e.g.

>ND3005_accessory_2122_length_3432

and an example of the terminal output is the following:

not found in FASTA file, returning empty sequence0_length_2460

Here are the commands I've been using:

$ samtools faidx Gene_db.fasta
$ xargs samtools faidx Gene_db.fasta < GH2_list.txt > GH2_output.fasta

Any thoughts would be appreciated. Thanks!

[finswimmer]

Hello spidcock01 ,

I'm not familiar with xargs. I would use parallel like this:

$ cat GH2_list.txt|parallel -k 'samtools faidx Gene_db.fasta {}' > GH2_output.fasta

And please use the formatting bar (especially the code option) to present your post better. I've done it for you this time.

Thank you!

fin swimmer

[spidcock01]

Unfortunately this still produces a file with just the headers. This means that the problem is probably with my list file, right?

Thank you for advice regarding formatting!

[finswimmer]

If you get just the headers, I would suspect that something with your fasta file is wrong.

What's the output of:

$ grep -A 10 "ND3005_accessory_2122_length_3432" Gene_db.fasta

Also you should check if the line ending terminators are correct:

$ file Gene_db.fasta
Gene_db.fasta: ASCII text

If the output is Gene_db.fasta: ASCII text, with CRLF line terminators you should install dos2unix and run dos2unix Gene_db.fasta.

fin swimmer

[h.mon]

Did you try my solution bellow?

[spidcock01]

Thanks for your help! You were right in that it was the line ending terminators.

Thank you again!

Answer 1

[h.mon]

samtools faidx doesn't take arguments from stdin, so you have to use a variable:

samtools faidx Gene_db.fasta $(cat GH2_list.txt) > GH2_output.fasta