Methods for nucleic acid amplification with thermostable ribonuc

Details Methods for nucleic acid amplification with thermostable ribonuc Methods for nucleic acid amplification with thermostable ribonuc

1997-07-15

1999-10-26

Degen, Nancy

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, 435 9121, 435 9151, 435183, 536 231, 536 243, 536 2431, C12Q 168, C12P 1934, C12N 900, C07H 2102

Patent

active

059726075

ABSTRACT:
Methods for nucleic acid amplification with thermostable ribonuclease H, in which single-stranded RNA (-) is prepared from RNA (+) as a target nucleic acid and the copy number of the single-stranded RNA (-) is increased through the use of thermostable ribonuclease H in combination with non-thermostable RNA-dependent DNA polymerase, non-thermostable DNA-dependent DNA polymerase and non-thermostable DNA-dependent RNA polymerase. In these methods, the number of amplification cycles is well increased and the sensitivity of detection can, therefore, be improved, as compared with the conventional methods. Also provided are methods for the detection of a target nucleic acid from RNA copies of a specific nucleic acid, obtained by any of the amplification methods, and reagent kits for use in these methods.

REFERENCES:
patent: 5409818 (1995-04-01), Davey et al.
patent: 5480784 (1996-01-01), Kacian et al.
Rapid and Simple Method for Purification of Nucleic Acids; Boom et al.; Journal of Clinical Microbiology Mar. 1990, p. 495-503, vol. 28, No. 3.
Preparation of Oligodeoxynucleotide-alkaline phosphatase conjugates and Their Use as Hybridization Probes Jablonski et al.; Nucleic Acids Research, vol. 14 No. 15, 1986, p. 6115-6128.
NASBA.TM. Isothermal Enzymatic In Vitro Nucleic Acid Amplification Optimized for the Diagnosis of HIV-Infection; Kievits et al.; Journal of Virological Methods, 35 (1991) p. 273-286.

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