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Team:Paris Saclay/Protocols/P1 phages transduction
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Revision as of 21:31, 20 August 2014
P1 phages transduction
Made overnight culture of the strains that you want to translate.
In the morning, add CaCl2 for a final concentration of 5mM.
In 4 tubes:
- Add 100µl phages culture in 0.1ml of bacterial culture.
- Add 10µl phages culture in 0.1ml of bacterial culture.
- Add 1µl phages culture in 0.1ml of bacterial culture.
- Negative control, just 0.1 of bacterial culture without phage.
Put at 37°C, during 20 to 30min.
Add 1ml of LB + Na citrate at 5mM final. Put at 37°C during 1 to 2 min.
Centrifuge and spread on LB + Na citrate at 5mM + antibiotic.
