Can PCR inhibitors act selectively to inhibit certain genes but allow others?
If you do your primer selection properly, why would you need to inhibit certain gene replication? If you have a specific enough primer, it should only lead to the replication of a specific gene. Use sequence searches to see if there any overlaps that are unwanted with your primer using this tool:
http://www.ncbi.nlm.nih.gov/tools/primer-blast/
Designing proper forward and reverse primers should lead to specific replication of a desired sequence.
Does that help at all?
No it may not be possible that inhibit or block one and other will be allowed. You may look the troubleshooting protocol in NEB support, it will help.
Firstly thanks for your concern but the situation that i conducted a multiplex PCR assay targeting 3 specific genes using 3 different primers but only one gene was detected in all tested samples so i am asking if PCR inhibitors can selectively inhibit the other 2 primers
It is possible, yes. I don't know much about PCR inhibitors in general, but I hear Exiqon Biofluids is much better than mirVana for getting rid of PCR inhbitors (this is according to the manufacturer claims).
I'm assuming you are doing RT-qPCR. If not, maybe the following:
http://www.promega.com/~/pdf/resources/articles/profiles-in-dna/2007/an-introduction-to-pcr-inhibitors/
Source:
http://www.exiqon.com/ls/Pages/news/biofluid-presentation.aspx
Witwer et al., 2013, DOI:10.4161/rna.25246
Read it in a paper that this protocol has better yields and better prevents accidental inhibition.
Hope this helps. As I've mentioned, I'm not an expert for accidental PCR inhibition.
Yes, differential inhibition can be a problem. See linked paper. We have also observed this and a multiplex reaction is of course somewhat more susceptible.
http://www.biomedcentral.com/content/pdf/1756-0500-1-70.pdf
Thanks Mr/Bustin this paper is important and useful for me
Sincerely yours
A.Oreiby
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