Index.Usage of PCR-Cycler

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1> Determine the annealing temperature according to the number -5 behind the TM(0.5M Na+) in the primer usage instructions. In order to prepare the concentrated solution, determine the amount of the ddH2O owing to the number *10 behind the nmol/OD.
2>Prepare the raw materials(50ul): buffer 5ul(not larger than 10%), 38ul of the ddH2O, 3ul of the Dntp3, 1ul of the previous primer, 1ul of the later primer, 1ul of the template, 1ul of the high fidelity enzyme, then make markers on the cap.
3>The normal process of the setting of the PCR-Cycler.
4>Determine whether the DNA suitability is successful or not.