"
Team:Paris Saclay/Protocols/P1 phages stock
From 2014.igem.org
(Difference between revisions)
(Created page with "=P1 phages stock= Made an overnight culture in which the phage have to grow. In the morning, add CaCl<sub>2</sub> at 5mM concentration final. In 4 tubes: #Add 10µl phages cul...") |
(→P1 phages stock) |
||
| Line 24: | Line 24: | ||
Recover the supernatant and stock it at 4°C. | Recover the supernatant and stock it at 4°C. | ||
| + | |||
| + | {{Team:Paris_Saclay/protocols_footer}} | ||
Revision as of 21:31, 20 August 2014
P1 phages stock
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.
