Have you tested the effect of inhibitors on PCR performance?
Yes. Please see Table 3 in our brochure Maximizing PCR and RT-PCR success. We tested the effects of different inhibitory substances in a number of PCR systems. We also analyzed the effect of including different volumes of reverse transcription (RT) reaction mixtures in PCR. Please see the table below for a list of commonly encountered template impurities and their inhibitory effects on PCR.
Impurities showing inhibitory effects on PCR
Substance | Inhibitory concentration |
SDS | >0.005% (w/v) |
Phenol | >0.2% (v/v) |
Ethanol | >1% (v/v) |
Isopropanol | >1% (v/v) |
Sodium Acetate | ≥5 mM |
Sodium Chloride | ≥25 nM |
EDTA | ≥0.5 mM |
Hemoglobin | ≥1 mg/ml |
Heparin | ≥0.15 i.U./ml |
Urea | >20 mM |
RT reaction mixture | ≥15% |