Wittrup: Site directed mutagenesis by QuikChange
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Site directed mutagenesis by QuikChange
Materials
- PfuTurbo(TM) DNA polymerase (2.5 U/uL)
- 10x reaction buffer
- Dpn I restriction enzyme (10 U/uL)
- dNTP mix
- Epicurian Coli(R) XL1-Blue supercompetent cells
- your primer 1
- your primer 2
- your plasmid
- Falcon(R) 2059 polypropylene tubes (15 mL)
- beads for top of reactions
- SOC Broth
- LB/Amp Plates
Protocol
Use protocol for Stratagene's Quikchange mutagenesis. Manual at: [1]
You will have to register to access this protocol.
Protocol notes:
- Add extra extension time to increase efficiency (1.5 to 2 min per kb). - Can do multiple QC's simultaneously by adding both primers to the PCR and sequencing more clones.
See other community protocols for this: Site-directed mutagenesis
