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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.

 

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