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Recombinant DNA Bacterial Transformation Student Instructions Mid-Log Culture Prep

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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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Presentation Transcript
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Recombinant DNA

Bacterial Transformation

Student Instructions

Mid-Log Culture Prep

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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.

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Recombinant Transformation

Day 2

Mid-Log Culture

Label a sterile 50 ml conical tube with your name and date.

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Recombinant Transformation

Mid-Log Culture

Use a sterile pipette to transfer one pipette full

(5 ml) of sterile LB broth into the conical tube.

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Recombinant Transformation

Mid-Log Culture

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.

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Recombinant Transformation

Mid-Log Culture

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.

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