This is a test version of Biostars. For the public version, visit https://www.biostars.org.
Is it possible to estimate the proportion of contamination from GC contents ?

Hello all,

I have multiple fastq files coming from different samples. Among them 2 show a significant diffrent GC content plot. I am wondering, is it possible from there to estimate the percentage of contaminated reads ?

Thanks

rna-seq

What do you mean by "a significant GC content plot"?

As genomax mentioned, no, you are not going to be able to determine this from GC content. If you have a large proportion of reads that don't map to the genome of your target organism, there are a few methods you could try.

I am doing de novo assembly. So far I am thinking of doing a pre-assembly with my samples with good gc content and then blast all my transcript to delete stranger transcripts. And then mapping all my reads to this transcriptom. And eventually do a final assembly with all mapping reads.

If you know/suspect that there is contamination, it may be best to address it up front before doing the assembly.

I have thougt about it but I can't blast 200 Gb of reads, I reduce considerably the data after assembly. Besides it seems to be a multi species contamination and I don't have the full genomes/transcriptomes of these associated species. So the other alternativethat was to identify the contaminants from a subset and then do a mapping on the full genome/transcriptom seem complicated.

Then you may want to treat your data as if it was a metagenomic dataset and use an assembler like metaSPAdes.

it possible from there to estimate the percentage of contaminated reads ?

I don't think so. You may get a hint that there is contamination e.g. with rRNA or a different species or something like that but you can't determine % of contaminant reads that may be present unless you go looking for those contaminant reads.

0 answers

No answers yet.

Log in to answer this question.