Isolation of plasmid for use in transformation

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Isolation of plasmid for use in transformation

Postby abclada » May 26 2017 9:11 pm

I did bacterial transformation using a GFP kit. Now I have green colonies growing on a petri dish. Can I isolate plasmids from my green colonies and use them to perform another transformation ? I thought I could but I tried with no success. I need your help
Thank you
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Re: Isolation of plasmid for use in transformation

Postby relaxin » May 30 2017 2:14 pm

First you need to pick one green colony and streak for single colonies. Then pick one single colony and isolate the plasmid DNA. You should be able to transform other competent cells. Since the plasmid is intact, even home-made low efficient competent cells should give you many colonies. Can you determine the yield of your plasmid DNA?
Retired academic researcher. Mention of a specific product does not imply my endorsement of the product. No conflict of interest or guarantee to work on the advice given. Do as I say, not as I do. Not liable to the loss of your valuable samples.
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