QuikChange Mutagenesis
Prepare PCR mixture .
Perform the PCR .
After PCR , add 1ul of DpnI to each pcr tube .
Incubate 1 hour - O / N at 37C .
PCR Purify using Qaigen kit .
Add 5ul of PCR reaction into 25ul DH5alpha or TG1 cells .
Incubate on ice for 5 minutes .
Heatshock at 42C for 45 seconds .
Recover on ice for 2 minutes .
Add 200ul LB ( if in 96 well or 8 strip format ) to each reaction .
Shake at 37C for 1 hour .
Plate on warmed antibiotic plates .
