Wittrup: Site directed mutagenesis by QuikChange: Difference between revisions
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==Site | ==Site directed mutagenesis by QuikChange== | ||
'''Materials''' | '''Materials''' | ||
| Line 23: | Line 23: | ||
'''Protocol''' | '''Protocol''' | ||
Use protocol for Stratagene's Quikchange mutagenesis. Manual at: [http://www.stratagene.com/products/showProduct.aspx?pid=573] | |||
See other community protocols for this | 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]] | |||
Latest revision as of 11:41, 2 July 2007
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
