( ing thermal cyclers with a heated lid ...)HotStarTaq PCR Handbook

ing thermal cyclers with a heated lid, do not use mineral oil. Proceed directly to step 6. Otherwise, overlay with approximately 50 l mineral oil.

6. Program the thermal cycler according to the manufacturers 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 and 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.

Troubleshooting Strategy

This troubleshooting strategy is a process that has been specifically designed to provide a convenient and time-saving optimization procedure for PCR using HotStarTaq DNA Polymerase or the HotStarTaq Master Mix Kit. It is recommended that the troubleshooting steps are followed in the order they are listed. The scientists in QIAGEN Technical Services are always happy to answer any questions you may have about either the information and protocols in this handbook or molecular biology applications (see inside front cover for contact information).

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