Amplification of mixed sequence nucleic acid fragments

Details Amplification of mixed sequence nucleic acid fragments Amplification of mixed sequence nucleic acid fragments

1991-08-02

2000-09-05

Marschel, Ardin H.

Chemistry: molecular biology and microbiology

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

Preparing compound containing saccharide radical

435 911, 435 915, 435 9151, 435 6, 436501, 536 231, 536 253, 935 7, 935 16, 935 17, 935 78, C12P 1934

Patent

active

061141496

ABSTRACT:
A method of amplifying a mixture of different-sequence DNA fragments which may be formed from RNA transcription, or derived from genomic single- or double-stranded DNA fragments. The fragments are treated with terminal deoxynucleotide transferase and a selected deoxynucleotide, to form a homopolymer tail at the 3' end of the anti-sense strands, and the sense strands are provided with a common 3'-end sequence. The fragments are mixed with a homopolymer primer which is homologous to the homopolymer tail of the anti-sense strands, and a defined-sequence primer which is homologous to the sense-strand common 3'-end sequence, with repeated cycles of fragment denaturation, annealing, and polymerization, to amplify the fragments. In one embodiment, the defined-sequence and homopolymer primers are the same, i.e., only one primer is used. The primers may contain selected restriction-site sequences, to provide directional restriction sites at the ends of the amplified fragments.

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