Friday, August 01, 2014

Who are the contaminants in your sequencing project? (crosspost from #microBEnet)

This was originally posted on microBEnet: Who are the contaminants in your sequencing project?

Well, been having many discussions recently about PCR amplification happening from "negative" controls where no sample DNA was added. Such amplification is alas pretty common - due to contamination occurring in some other material added to the PCR reaction.  Obviously it would be best to eliminate all DNA contamination of all reagents and all PCRs.  But if that does not happen, it is possible to try to detect contamination after it has happened.  Below I post some papers related to post-sequencing detection of contamination:
Any other suggestions or comments would be welcome. UPDATE 10:30 AM 7/25 - Was reminded on Twitter of a new, critically relevant publication on this issue: Reagent contamination can critically impact sequence-based microbiome analyses

1 comment:

  1. 2nd try:-(
    A huge but so far little regarded aspect is not contamination of samples or of physical substances but of computer software and databases.
    We have found Mycoplasma genes in a variety of Bioinformatics tools. There are even Mycoplasma genes in the reference human genome at NCBI BioTechniques, Vol. 47, No. 6, December 2009 pubmed/20047202

    In Silico Infection of the Human Genome

    More Mouldy Data: Another mycoplasma gene jumps the silicon barrier into the human genome

    Sneaky Bacteria Impersonate as Humans

    There is even contamination in the other direction with Human genetic sequences appearing in online publicly accessible plant genomes: Longo 2011