De novo polynucleotide synthesis using rolling templates

Details De novo polynucleotide synthesis using rolling templates De novo polynucleotide synthesis using rolling templates

1997-09-15

2000-10-24

Riley, Jezia

Chemistry: molecular biology and microbiology

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

Preparing compound containing saccharide radical

435 6, 435 912, 435810, 536 221, 536 243, 536 231, 536 2431, 536 253, C12Q 168, C12P 1934, C07H 1900, C07H 2104

Patent

active

061365688

ABSTRACT:
Disclosed is a method for synthesizing polynucleotide molecules such as genes or gene segments. A primer having 5' and 3' ends is incubated with a relatively shorter template having a 5' region non-complementary to the primer, a 3' region complementary to the 3' end of the primer, and a non-reactive 3' terminus to allow the 3' region of the template to anneal to the primer. The annealed product is reacted with at least one nucleotide in the presence of a template-dependent polynucleotide polymerase to produce a primer extended at its 3' end by at least one nucleotide complementary to the 5' region of the template. The extended primer is then dissociated from the template. The extended primer is further extended by repeating this cycle for sufficient cycles, wherein the templates and enzymes may differ from cycle to cycle, to obtain the object polynucleotide. Also disclosed are template libraries and kits containing said libraries for use in conjunction with the polynucleotide synthesis method.

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