running a gel on a colony - basic question (Dec/04/2008 )
I streaked transformed vector DNA onto LB plates by using electrocompetent cells to transform. I have a lot of great colonies. Now I want to run a gel to look at the DNA. How do I isolate the DNA from the cell to look at it on the gel? Can someone provide me a protocol? I think I am supposed to use phenol/chloroform but not sure. Thanks.
Do you want to assess the quality of the DNA, or make sure they transformed properly? You could do a colony PCR and run that on a gel.
Or do you actually want to extract the DNA from the colonies?
Do you want to assess the quality of the DNA, or make sure they transformed properly? You could do a colony PCR and run that on a gel.
Or do you actually want to extract the DNA from the colonies?
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Yes, exactly my question as well.
To isolate DNA from the colonies (to check integrity, e.g.) you should grow them in rich media overnight and do a genomic DNA isolation (many protocols available).
If you have primers specific for the plasmid you transformed with, you can resuspend a teeny bit of the colony in ~100uL water (assuming it is E. coli), boil, and use 1-2uL as a template in a PCR reaction with said primers to check the presence of the plasmid.