Phi80 Integration Protocol

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  • 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