FCCT Biochemistry Lab:PCR protocol iGEM HS

iGEM Protocols

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Protocols PCR Amplification PCR with Taq polymerase Setup Amplification PCR Reaction 1 μL Template DNA 11 μl deionized (dI) H2O 1 μl 10μM forward primer 1 μl 10μM reverse Primer 2 μl 10x Taq Polymerase Buffer 2 μl MgCl2 1 μl 10x dilluted taq polymerase Total 20μl Amplification PCR Reaction Program 1. 95 °C for 5min 2. 95 °C for 30s 3. 50 °C for 90s (important step) 4. 72 °C for 1min per kb target gene plus 10-15% (important step) 5. Repeat 2-4 25x 6. 72 °C for 5 to 10min 7. 4 °C for forever (don’t forget to turn the machine off the next day)