Methods:M13-mediated phagemid transduction
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Transfer of plasmids without transformation. Plasmids must contain a compatible origin of replication
- M13 rv-1 helper phage
- phagemid-containing cells (e.g. from pZ150 or pZ152)
- 2X YT media
- prepare phage stock as in Methods:CTAB_M13_phagemid_prep
- pasteurize the phage stock by heating at 55-60°C for 15-20 minutes
- use 5 μl phage with 50 μl of a fresh overnight of a susceptible host
- incubate at 37 degrees C for 30 minutes to allow expression of antibiotic resistance genes
- plate on agar with appropriate antibiotics
This method gives high enough transduction so that you can spot a few microliters of the transduction mix on the plate and streak for single colonies from the spot.
M13 transduction is somewhat mutagenic. Mutations in lambda cI plasmids occurred at approx 10-2 - 10-3 per transductant.
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