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Wednesday, May 1, 2013

Restriction Digestion


Restriction Digestion

This is an explanation for performing Restriction Digestion using restriction digestion enzymes, some of the examples of Restriction enzymes are E.coRI, BamHI, SalI, etc. each of these enzymes can specifically recognize particular regions of the target DNA and cleave that region to give fragments of different lengths. Single and Double restriction digestions are usually performed for molecular biology cloning related works. there are various restriction digestion enzymes are available suitable one need to selected depending on the target sequence and the product required.
Restriction digestion reaction volume will be usually 50uL, containing the target DNA, restriction enzyme and the assay buffer. The reaction mixture is incubated for 1hr at 37oC, an agarose gel electrophoresis is performed after the incubation period to check the restricted fragments. it is recommended to have a control reaction with out any enzyme. while performing agarose gel electrophoresis don’t forget to load molecular weight marker for comparison of molecular weight of  the restricted fragments.
Restriction digestion