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Team:Paris Saclay/Protocols/P1 phages stock
From 2014.igem.org
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Made an overnight culture in which the phage have to grow. | Made an overnight culture in which the phage have to grow. | ||
Revision as of 21:38, 20 August 2014
P1 phages stock
a
b
Made an overnight culture in which the phage have to grow.
In the morning, add CaCl2 at 5mM concentration final.
In 4 tubes:
- Add 10µl phages culture in 0.1ml of bacterial culture.
- Add 50µl phages culture in 0.1ml of bacterial culture.
- Add 100µl phages culture in 0.1ml of bacterial culture.
- Negative control, just 0.1 of bacterial culture without phage.
Leave 10 minutes at room temperature.
Add 1ml LB in each tube. Add 3ml Agarose 6g/l (at 65°C).
Spread on cold LCA dish + CaCl2 at 5mM final.
Incubate during 5 to 7hours at 37°C, or during the night at 30°C.
When the carpet becomes limpid, harvest it with a rake in a Falcon tube.
Add 0.5ml of chloroform, and centrifuge 10min at 5000g.
Recover the supernatant and stock it at 4°C.
