Gel Electrophoresis and PCR troubleshooting

Check the concentration of template DNA, as high concentrations can inhibit PCR amplification. Dilute if necessary. Set a higher annealing temperature and reduce extension time to avoid non-specific binding and amplification. Use only 0.5u of Taq polymerase (instead of 5u), and carefully dilute the first round product (1:1000) for the second round PCR, running 20-25 cycles to prevent over-amplification. https://www.youtube.com/watch?v=kUbrM-cAk34

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