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  • 發布時間:2019-10-17 21:39 原文鏈接: PCR基本實驗方法(二)

    Recommended Reaction Conditions:

    Initial Conditions:

    Initial denaturation at start: 92 - 97oC for 3 - 5 min. If you denature at 97oC, denature sample only; add rest of mix after reaction colls to annealing temperature (prevents premature denaturation of enzyme).

    Initial annealing temperature: as high as feasible for 3 min (eg: 50 - 75oC). Stringent initial conditions mean less non-specific product, especially when amplifying from eukaryotic genomic DNA.

    Initial elongation temperature: 72oC for 3 - 5 min. This allows complete elongation of product on rare templates.


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