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HotStarTaq PCR Handbook

x).

5. When using a thermal cycler with a heated lid, do not use mineral oil. Proceed directly to step 6. Otherwise, overlay with approximately 100 l mineral oil.

6. Program the thermal cycler according to the manufacturer's instructions.Each PCR program must start with an initial heat activation step at 95C for 15 min.A typical PCR cycling program is outlined below. For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.



7. Place the PCR tubes in the thermal cycler and start the cycling program.

Note: After amplification, samples can be stored overnight at 28C or at 20C for longer storage.

PCR Protocol Using HotStarTaq DNA Polymerase and Q-Solution

This protocol is designed for using Q-Solution in PCR assays. Q-Solution changes the melting behavior of DNA and can be used for PCR systems that do not work well under standard conditions. When using Q-Solution the first time for a particular primertemplate pair, always perform parallel reactions with and without Q-Solution. This recommendation should also be followed if another PCR additive (such as DMSO) was previously used for a particular primertemplate pair.

When using Q-Solution, the following effects may be observed depending on the individual PCR assay:

Case A: Q-Sol
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