Method for producing a polynucleotide for use in single primer a

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

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435 6, 935 77, 935 78, C12P 1934, C12Q 168

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055958915

ABSTRACT:
A method is disclosed for producing a single stranded polydeoxynucleotide having two segments that are non-contiguous and complementary with each other. The method comprises the steps of providing in combination (1) a polynucleotide having two non-contiguous, non-complementary nucleotide sequences S1 and S2 wherein S2 is 5' of S1 and is at least ten deoxynucleotides long and (2) an extender probe comprised of two deoxynucleotide sequences, wherein the sequence at the 3'-end of the extender probe is hybridizable with S1 and the other of the deoxynucleotide sequences is homologous to S2 and (b) extending the extender probe along the polynucleotide. The method can also comprise providing in the combination a polydoxynucleotide primer capable of hybridizing at least at its 3'-end with a nucleotide sequence complementary to S2 under conditions where (1) the extended extender probe is rendered single stranded, (2) the polydeoxynucleotide primer hybridizes with and is extended along the extended extender probe to form a duplex comprising extended primer, (3) the extended primer is dissociated from the duplex, and (4) the primer hybridizes with and is extended along the extended primer to form a duplex comprising extended primer, and repeating steps (3) and (4). The method finds particular application in the detection of polynucleotide analytes.

REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
patent: 5066584 (1991-11-01), Gyllensten et al.
patent: 5508178 (1996-04-01), Rose et al.
Frohman et al., PNAS(USA)85:8998-9002 (Dec. 1988).
Gullensten et al., PNAS(USA)85:7652-7656 (Oct. 1988).
Nelson et al., PNAS(USA)86:6686-6690 (Sep. 1989).

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