Recombinant DNA Bacterial Transformation Student Instructions Mid-Log Culture Prep. Recombinant Transformation Day 1 Overnight Culture Plate. Using the extra “LB” agar plate, streak the MM 294 E.coli culture on the plate and incubate 18-24 hours. Recombinant Transformation Day 2
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Mid-Log Culture Prep
Overnight Culture Plate
Using the extra “LB” agar plate, streak the MM 294 E.coli culture on the plate and incubate 18-24 hours.
Label a sterile 50 ml conical tube with your name and date.
Use a sterile pipette to transfer one pipette full
(5 ml) of sterile LB broth into the conical tube.
Locate a well-formed colony from your plate of bacteria grown the day before. Use a sterile inoculating loop, bent paper clip, or toothpick to scrape up the colony and transfer it to the LB broth in the conical tube. Your instructor may have started the culture several hours before class.
Incubate the tube for 3-4 in a shaking water bath or use an
orbital shaker in an incubator.
Alternatively: Incubate 4 hours in the incubator with occasional shaking
by hand. For best results, shake the tube a couple of times an hour
when possible. After the 4 hour incubation period, immediately
proceed to the “Making Competent Cells” protocol.