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查看更多产品信息 10966-018 - FAQs (10966-018)
3 个常见问题解答
Here are some suggestions for optimizing your PCR under such conditions:
-Make sure primers don't have complementary sequences at the 3' ends.
-Optimize the annealing step by increasing the temperature in 2-5 degrees C increments, and minimizing the annealing time. You can try higher annealing temperatures in the first few cycles and lower annealing temperatures in the subsequent cycles.
-Try hot-start protocols.
-Optimize the magnesium concentration for each template and primer combination.
-To minimize chances of amplifying contaminating DNA, use aerosol-resistant tips and UDG.
Platinum Taq DNA Polymerase can be used in the same manner as standard Taq DNA polymerase. One unit of Platinum Taq DNA Polymerase incorporates 10 nmol of deoxyribonucleotide into acid-precipitable material in 30 min at 74 degrees C.
1 unit per reaction is sufficient for most reactions/applications, but in some instances it may be necessary to use more enzyme.
NOTE: the number of reactions specified on vials of Life Technologies polymerases assumes you are using 1 unit per reaction.
Platinum Taq DNA Polymerase is designed for hot-start PCR. Because it is bound by a monoclonal antibody, Platinum Taq DNA Polymerase is inactive when the temperature is below 94 degrees C. The high-temperature step at the beginning of the PCR cycle (typically 94 degrees C) denatures and dissociates the antibody from the polymerase, restoring it to full activity.
Mispriming typically occurs at the start of PCR. Using a hot-start polymerase such as Platinum Taq DNA Polymerase dramatically reduces mispriming, as it has little to no activity until after the denaturation step. Therefore, hot-start PCR--mediated by Platinum Taq DNA Polymerase--has a higher specificity than PCR with standard Taq DNA Polymerase.