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-   -   Amplicon sequencing remove primer & adapters (http://seqanswers.com/forums/showthread.php?t=21767)

wzhang1001 07-17-2012 11:03 AM

Amplicon sequencing remove primer & adapters
 
Hi,

I have amplicon sequencing experiment with amplicon length between 45 ~124 bases. The sequencing length is 150, which completely covers the amplicon.

I wanted to remove the primers and adapters in the fastq file, which seems to be a little challenging becase we don't know the sequence of primers (but we do know the adapter sequence).

But we know the sequence of amplicons, so I'm thinking if there is any way that I can retrieve the sequence of amplicons from the the fastQ file, which will automatically get rid of both primer and adapters. Of course I need also retrieve the correspnding Q score for the amplicon sequences.

Is there any tool I can use for this purpose?

Thanks in advance for your help!

JackieBadger 07-17-2012 12:38 PM

You can use PRINSEQ http://edwards.sdsu.edu/cgi-bin/prin...seq.cgi?home=1
As far as I remember you can tell the program to trim a certain amount of bases from either end.
You can also do this with Galaxy: https://main.g2.bx.psu.edu/

Prinseq may be better as you will retain your Q data.

If you wanted to sort your amplicons while you remove primers and adapters you could use jMHC http://code.google.com/p/jmhc/
For this you need to know your primer sequence but you should be able to figure out what your primers were from your sequence.

jlli 07-18-2012 05:17 AM

You can try

cutadapt : http://code.google.com/p/cutadapt/
or
fastx clipper : http://hannonlab.cshl.edu/fastx_toolkit/index.html

Tha cutadapt has more options.

Krish_143 07-18-2012 05:23 AM

You can also try

Trimmomatic : http://www.usadellab.org/cms/index.php?page=trimmomatic

wzhang1001 07-19-2012 10:32 AM

Hi all,

Thank youf or all your help!


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