LAB - pGLO Restriction Digest

pGLO plasmid map
click on picture for a full size image

Following the plasmid isolation procedure using minipreps, our next step would be the one we would follow if we were cloning a gene: Isolate the gene from the plasmid. In this case we want to isolate the GFP gene, and we will be using the BamH I restriction enzyme, which cuts GFP from the plasmid, and it also cuts the gene in two fragments.

First we ran an agarose gel to know if we actually have succesfully isolated our plasmid DNA (we do know we have plasmid DNA in some of our section 2 samples, but it is still unclear in our section 1 samples). After that, we set up the restriction enzyme digest (RED).

Next week we will run a polyacrylamyde gel (PAGE) that we will silver stain. Silver staining is a very sensitive technique to visualize proteins and DNA in agarose and polyacrylamide gels.

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