FAQs
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FAQs
Browse our frequently asked technical questions by selecting a product below.
Why is there a non-specific product when using the same primers and PCR conditions as a competitor’s product?
It’s most likely because the time for the 1st step (hot start) is too short. Ensure that the hot start phase is done at 95°C for 2 minutes to fully activate the enzyme. The recommended thermal profile is:
- 95°C (120 seconds)
- 40 cycles: 95°C (5-15 seconds) – 60°C (20-30 seconds)
- Melt
If non-specific products are still obtained, we recommend raising the annealing/extension temperature from 60°C to 65°C, depending on the primer set used.