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DNA Endonuclease digestion

DNA Endonuclease digestion - DNA Techniques

DNA Endonuclease digestion - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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Old 03-08-2012, 03:44 PM
Pipette Filler
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Default DNA Endonuclease digestion



I am currently trying to insert a gene of interest from a plasmid into a different empty plasmid. In order to cut the gene of interest from the plasmid, I know I have to use restriction enzymes. However, since the plasmid is circular, would I have to cut the plasmid with two different restriction enzymes or one restriction enzyme that cuts at two places? Also, if the restriction enzyme is supposed to match the enzyme I will be using for the empty plasmid, how is this possible if I use two different restriction enzymes for the plasmid containing my gene of interest?

I'm just confused about the details of this procedure because all of the research I have found online relates to a gene of interest that is not in a plasmid.

If anyone could help me, that would be great. Thanks!
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