I am investigating differential expression of specific functional genes of bacteria from environmental RNA samples (marine sediment) and I am looking for control genes to normalise my samples. I found that most people use either 16s rRNA or rpoB but I would like to know if other conserved genes with primers targeting most bacteria have been developed. The MIQE guidelines for RT-qPCR recommend using at least 3 control genes.
Hi,
you can find solution here: https://www.ncbi.nlm.nih.gov/pubmed/26149127
Good luck!
Thank you Marcela. I had read this paper and found it very interesting but realised that for most of the suggested genes, the developed primers are targeting specific species, genera, etc. I'm just wondering if someone has developed a set of primers (for any control genes) that targets most bacteria. I found one paper where researchers developed such a primer set with the tuf gene (see ref. below), but this is the only one I found so far.
Tanaka, Y., Takahashi, H., Simidu, U., 2010. Design of a new universal Real-Time PCR system targeting the tuf gene for the enumeration of bacterial counts in food. J. Food Prot. 73, 670–679.
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