Difference between revisions of "Phi80 Integration Protocol"
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*swab the plate and outgrow in 5mL LB+gentamicin to saturation | *swab the plate and outgrow in 5mL LB+gentamicin to saturation | ||
*restreak on LB+gentamamicin at 37 degrees | *restreak on LB+gentamamicin at 37 degrees | ||
| − | + | Note: Protocol adapted from Anderson Lab, UC Berkeley | |
Latest revision as of 18:07, 18 September 2008
- Prepare chemically competent cells of the strain you want to integrate into
- Transform in pInt80-649, plate on amp at 30 degrees
- Pick a single colony, grow a starter ON
- Dilute 200uL to 5mL, grow at 30 degree to mid log
- Do another chemically competent cell prep
- Transform in the phi80 CRIM plasmid you wish to integrate
- Rescue 1 hr in LB, plate on gentamicin at 37
- swab the plate and outgrow in 5mL LB+gentamicin to saturation
- restreak on LB+gentamamicin at 37 degrees
Note: Protocol adapted from Anderson Lab, UC Berkeley
