Melbourne/Site directed mutagenesis


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  • Application: Single point mutation in plasmid exceeding 8Kbp
  • Time to complete protocol from DNA template:3 days (including 2 overnight incubations)
Lab time: waiting time method
2 hours (set up PCR) 4 hours (PCR) stratagene kit manual
30min (add dnp) 1 hour(digest) stratagene kit manual
1 hour (transformation) 1 hour(NZY media) stratagene kit manual
1 hour (plate out) 16 hours (grow overnight)
1 hour (pick) 16 hours (culture overnight)
3 hours (miniprep,and glycerol stock) Wizard kit manual -->next mutation cycle-->
1 hour (set up Digest, make gell) 2 hours
1/2 hour (Load Gell) 2 hours
1/2 hour (Photo)
  • Approximate cost of materials for 10 mutations: AUD $2,000.00

Primary & secondary Reagents Required including controls
Equipement Required
  • PCR machine
  • microcentrifuge: 0.6ml
  • ice
  • waterbath at 42 deg C
  • 37 deg C incubator with shaker
  • gel manufacture and elecctrophoresis equipment for confirmation.
  • Camera
Method including controls

  • PhillipDodson 03:48, 1 July 2007 (EDT):Secondary Reagent Template 1/July/2007