Coupled amplification and sequencing of DNA

Details Coupled amplification and sequencing of DNA Coupled amplification and sequencing of DNA

1993-07-28

1995-06-27

Fleisher, Mindy B.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, 536 2433, C12Q 168, C12P 1934, C07H 2104

Patent

active

054279119

ABSTRACT:
A process for sequencing DNA segments having complementary strands comprising (a) synthesizing simultaneously truncated products and full-length products starting from both 3' ends of the complementary strands of the DNA segment, which serves as a template, by introducing specific oligonucleotide primers annealing to the 3' ends of both complementary strands of the DNA segment, a deoxyribonucleotide elongator for each of adenine, guanine, cytosine and thymine, a thermally stable DNA polymerase and a terminator for each of adenine, guanine, cytosine and thymine, (b) thermally cycling step (a), (c) separating out the resultant truncated products and full length products according to size, and (d) selectively detecting all truncated products and full length products according to size, and (e) selectively detecting all truncated products and full-length products generated from a given strand of template.

REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
K. B. Mullis and F. A. Foloona, "Specific Synthesis of DNA In Vitro via a Polymeric-Catalyzed Chain Reaction," Methods in Enzymology, 155, 335-350.
U. B. Gyllensten and H. A. Erlich, "Generation of Single-Stranded DNA by the Polymerase Chain Reaction and Its Application to Direct Sequencing of the HLA-DQA Locus," PNAS USA, 85, 7652-7656 (1988).
Ruano and Kidd, Proc. Natl. Acad. Sci. USA, Apr. 1991, v. 88, 2815-19.
Ruano and Kidd, Nucleic Acids Res, v. 19, 1991, pp. 6877-6882.
Carethers et al, BioTechniques, vol. 7, May 1989, pp. 494-498.
Nurakawa et al, DNA, v. 7, 1988, pp. 287-295.
Sarger et al, Proc. Natl. Acad. Sci USA, vol. 74, No. 12, Dec. 1977, pp. 5463-5467.

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