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    Step 5: Isolation of colonies with Spectinomycin Resistance and sequencing
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    Step 5: Isolation of colonies with Spectinomycin Resistance and sequencing

    Saturday, 2/22/2020Step5.jpg
    Saturday, 2/22/2020
    1.
    Plate the rest of the transformation onto ~10 Agar plates prepared with both Ampicillin (or Carbenicillin) and Spectinomycin.
    2.
    Incubate at 37ºC for ~12-18h (I incubated them and check at 20 and 46h time). Any colonies that grow should contain the antibiotic resistance marker you isolated from the DNA library! Congratulations! Circle one or two of these colonies and send them to Genewiz for Sanger sequencing. Wrap the plate in parafilm.
    Results
    +control: Joe and I did it together, we did not get any colonies in any of the plates or liquid cultures. No spark on the electroporator either so not sure what happened
    -control: Joe and I did it together, we got colonies in the carb plates and the liquid cultures (both carb and carb+spec) but not in the carb + spec plates. This was quite weird, they were just cells for which we did not do anything, just add the SOC media.
    Transformation: I had clear colonies on some of my carb plates and none for the carb+spec but Joe got some in the carb+spec!. No colonies either in the liquid cultures.
    Step5.jpg
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