I have DNA samples that have been extracted from stool samples, which I am running qpcr on for diagnostics. I was wondering what to use as an internal amplification control, since stool has a lot of pcr inhibitors and is a complex matrix so I couldn't think of what human gene would be present in a consistent amount.
I have primers and probes for phocine herpes virus (PhHV), and was wondering if I could spike the DNA samples with a gene fragment from PhHV as an internal control?
Hi, in principle this would work but be aware that you need to highly dilute this as you would need to come to ~1000 copies/ reaction. You might also look for options some suppliers do offer already as you also need to make the IC assay more susceptible for inhibition than your target assay
Sven
I could recommend the expression of genes active in colonic epithelial cells. They are frequent in stool samples, and the expression of housekeeping genes should be informative. The problem is only the number of cells in each stool sample, but I think it can be calculated as an average number of cells.
- Badges
- Science method