Method of double stranded DNA synthesis

Details Method of double stranded DNA synthesis Method of double stranded DNA synthesis

1997-07-22

2000-11-07

Guzo, David

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 911, 435 915, 435 9152, 536 231, 536 253, C12P 1934, C07H 2104

Patent

active

061435317

ABSTRACT:
The present invention provides an improved method for the synthesis of double stranded DNA, particularly complementary DNA for the construction of directional complementary DNA libraries. The method comprises synthesizing a first strand of DNA complementary to a selected RNA or DNA template by contacting with the template a linker/primer comprising a selected restriction site, a suitable RNA or DNA dependent DNA polymerase, and substrates comprising a deoxyribonucleotide triphosphate analog. The linker/primer and deoxyribonucleotide triphosphate analog are selected such that incorporation of the nucleotide analog in the first strand substantially protects the double stranded DNA from cleavage, under conditions sufficient to cleave or substantially cleave the linker/primer, at the selected restriction site.

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