PCR Internal Controls - (Jun/13/2006 )
I'm doing a multiplex PCR for the amplification of chlamydia and gonorrhoea and really want an endogenous internal control as I'm using urine samples that contain up to 25% inhibitors. I'm very new to this field so can anyone please suggest genes that may be suitable for internal control purposes?
thank you so much!!!
There will almost certainly be some bacteria. You could use the 16S ribosomal universal primers to amplify whatever bacterial DNA is around.
IMHO, if you want an endogenous control for urine samples, don't use bacteria
some of these patients may have been affected by recent antibiotic use (not necessarily correct ones for CT and GC); not all our flora is the same, too
I would pick a gene from a protein expressed in the epithelial cells lining the urinary tract ( one of the standard HKG's). those are going to be in the samples anyways, if you are testing for GC