This is a test version of Biostars. For the public version, visit https://www.biostars.org.
extracting certain region 16sRNA from Silva

I would like to extract V4 region from all available sequences on Silva; If I am not mistaken, Silva is a 16sRNA database, but it does not have such annotate on its 16sRNA sequences.

16srna sequencing

3 answers

Looking here you will see the location and structure of the V4 region

http://www.nature.com/nrmicro/journal/v12/n9/fig_tab/nrmicro3330_F1.html

Then Import Silva db in ARB, open the alignment editor and there you should be able to recover that region. At the moment my ARB is not working, so I cannot give u more details. But maybe you could give it a try and figure it out by your self the right way of doing it.

I believe the one you sent is only for Escherichia coli,

how about other species ?! I don't think, they would necessarily even be with the same length

The alignment Silva and ARB are using refer to E. coli. When you submit your seqs for alignment they do not perform a de-novo alignment but align you seqs on the reference. That's why I think it would make sense to use ARB to take the reference position

o - cool - does that mean, I should download the aligned file; or the pure fasta file and align it against E. coli and ... ? Thanks

I thought you could just download the silva, load it into ARB and use the alignment editor to extract what u need.

BBMap contains tools for this purpose, based on primer sequences. Let's say you want the region between primers "ACGT" and "CCCC":

msa.sh in=16s.fa out=x.sam literal=ACGT
msa.sh in=16s.fa out=y.sam literal=CCCC
cutprimers.sh in=16s.fa out=V4.fa sam1=x.sam sam2=y.sam

If you have multiple primers, use a comma-delimited list such as literal=ACGT,ACTT.

I've recently developed a simple Python tool to extract 16S regions with respect to the reference E. coli notation (V4 is mapped from ~515 to ~810). The tool is openly available here https://github.com/grayfall/hypervar. You can install it using pip: pip install git+https://github.com/grayfall/hypervar (I recommend doing it in a virtualenv environment). The installation script adds extract.py executable to your PATH, hence it can be invoked as extract.py -f your_seqs.fna -o extracted.fna -s 515 -e 810 -j cores. I strongly recommend using multiple cores if you are willing to process many thousands of sequences, because the tool uses relatively slow BioPython alignment functions.

Log in to answer this question.