Wittrup: Site directed mutagenesis by QuikChange

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:

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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