Restriction selection cloning: a simple general method for the selection of recombinant DNA.
Shelley J. Russek, Jeremy Quirk, David H. Farb
Abstract
Shelley J. Russek, Jeremy Quirk, David H. Farb
Abstract
It is well known that the ligation of two DNA fragments which are the product of digestion from different restriction enzymes will not lead to the regeneration of either of the original blunt end or cohesive end restriction sites. This property of sequence incompatibility for the original restriction enzyme can be exploited in a general cloning procedure for both PCR products and restricted DNAs. Restriction selection is particularly useful when cloning low abundance polymerase chain reaction (PCR) products and when cloning blunt ended DNA into reporter vectors that lack a method for the selection of recombinants.
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It is well known that the ligation of two DNA fragments which are the product of digestion from different restriction enzymes will not lead to the regeneration of either of the original blunt end or cohesive end restriction sites. This property of sequence incompatibility for the original restriction enzyme can be exploited in a general cloning procedure for both PCR products and restricted DNAs. Restriction selection is particularly useful when cloning low abundance polymerase chain reaction (PCR) products and when cloning blunt ended DNA into reporter vectors that lack a method for the selection of recombinants.
Key concepts: Restriction enzyme, Cloning (programming), Recombinant DNA, Ligation, Restriction site, Restriction digest, Sticky and blunt ends, Molecular cloning