Direct cloning of PCR amplified nucleic acids

Details Direct cloning of PCR amplified nucleic acids Direct cloning of PCR amplified nucleic acids

1993-09-09

1996-01-30

Jones, W. Gary

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

435 9141, 435 912, 4353201, C12P 1934

Patent

active

054879930

ABSTRACT:
Methods are described for producing recombinant DNA molecules from suitable host vectors and nucleic acids subjected to 3'-terminal transferase activity. In one embodiment, the method takes advantage of the single 3'-deoxy-adenosine monophosphate (dAMP) residues attached to the 3' termini of PCR generated nucleic acids. Vectors are prepared with recognition sequences that afford single 3'-terminal deoxy-thymidine monophosphate (dTMP) residues upon reaction with a suitable restriction enzyme. Thus, PCR generated copies of genes can be directly cloned into the vectors without need for preparing primers having suitable restriction sites therein. The invention also contemplates associated plasmid vectors and kits for implementing the methods.

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