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FAQs
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What are the recommendations for diluting UltraScript® 2.0 to avoid RTase-induced Taq inhibition?
- Diluting UltraScript® 2.0 prior reverse transcription.
- Assuming 2.5 µL of RTase reaction is transferred to 20 µL of PCR reaction (4x dilution)
- We recommend diluting the RTase 10x – 50x to nullify the effects of RTase-induced Taq inhibition.
- We recommend the following buffer or equivalent for the dilutions:
- 20 mM Tris/Cl pH 8.0 (25°C)
- 100 mM KCl
- 1 mM EDTA
- 1 mM DTT
- 5 % (w/v) Tween 20
- 5% (w/v) Triton X-100
- 50% (v/v) Glycerol
- Diluting UltraScript® 2.0 after reverse transcription, that is, diluting the reaction mixture
- Diluting the RT reaction mixture dilutes out the inhibitory effect of the RTase as well as cDNA therefore we recommend dilutions between 5x – 15x to find the optimal result.
- Assuming 2.5 µL of RTase reaction is transferred to 20 µL of PCR/qPCR reaction, 4x dilution for a total of 20x – 60 x. Use water for RT reaction dilutions.
