Transform pCas9
Qiaowa Gong
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Abstract
this protocol is aimed to provide instructions of transforming pCas9, which is a key step of knock-out gene by Crispr/Cas9
Steps
Mix 50µL of competent MG1655 with 10µL of pCas9
Keep the reaction mix on ice for0h 30m 0s
Then, transfer the mix to the water bath set at 42°C for 0h 0m 45s
Next, keep the mix on the ice for 0h 2m 0s
Add 940µL LB broth and incubate the cells at 37°C with vigorous shaking (200 rpm) for 1h 0m 0s
Plate the cells on LB-agar plates containing chloramphenicol
Incubate at 37°C
