Direct cloning of PCR amplified nucleic acids

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

1996-07-18

1998-10-27

Houtteman, Scott W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

536 231, C12Q 168

Patent

active

058276572

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.

REFERENCES:
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Denney, et al. (1990) "DNA Generated by Polymerase Chain Reaction Using Taq DNA Polymerase has Non-Template Nucleotide Additions: Implications for Cloning PCR Products", Amplification. A Forum for PCR Users, No. 4, The Perkin Elmer Corporation US, pp. 25-26.
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