Random-primed reverse transcriptase- in vitro transcription...

Details Random-primed reverse transcriptase- in vitro transcription... Random-primed reverse transcriptase- in vitro transcription...

2007-06-12

2007-06-12

Horlick, Kenneth R. (Department: 1637)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

Reexamination Certificate

active

10432176

ABSTRACT:
A random-primed reverse transcriptase-in vitro transcription method of linearly amplifying RNA is provided. According to the methods of the invention, source RNA (or other single-stranded nucleic acid), preferably, mRNA, is converted to double-stranded cDNA using two random primers, one of which comprises a RNA polymerase promoter sequence (“promoter-primer”), to yield a double-stranded cDNA that comprises a RNA polymerase promoter that is recognized by a RNA polymerase. Preferably, the primer for first-strand cDNA synthesis is a promoter-primer and the primer for second-strand cDNA synthesis is not a promoter-primer. The double-stranded cDNA is then transcribed into RNA by the RNA polymerase, optimally in the presence of a reverse transcriptase that is rendered incapable of RNA-dependent DNA polymerase activity during this transcription step. The subject methods produce linearly amplified RNA with little or no 3′ bias in the sequences of the nucleic acid population amplified.

REFERENCES:
patent: 5554516 (1996-09-01), Kacian et al.
patent: 5716785 (1998-02-01), Van Gelder et al.
patent: 5932451 (1999-08-01), Wang et al.
patent: 6132997 (2000-10-01), Shannon
patent: 6235483 (2001-05-01), Wolber et al.
patent: 6495320 (2002-12-01), Lockhart et al.
patent: WO 01/09310 (2001-02-01), None
patent: WO 01/71036 (2001-09-01), None
Ahern, H., “Biochemical, Reagent Kits Offer Scientists Good Return on Investment,”The Scientist 9(15):20-24, 1995.

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