Title:
METHOD FOR SYNTHESIZING COMPLEMENTARY DNA
Document Type and Number:
WIPO Patent Application WO/2014/061819
Kind Code:
A1
Abstract:
Provided is a method which enables the highly efficient reverse transcription of a double-stranded RNA virus and also enables the amplification of the resultant reverse transcript. A method for synthesizing complementary DNA using double-stranded RNA as a template, said method being characterized in that a mixed solution of double-stranded RNA extracted from a sample and a primer is subjected to steps (1) and (2) as mentioned below in this order, and subsequently single-stranded DNA is synthesized from the mixed solution using a reverse transcriptase: (1) a step of heating the mixed solution at a temperature equal to or higher than 95˚C and then rapidly cooling the mixed solution; and (2) a step of heating the mixed solution at a temperature lower than 90˚C and then rapidly cooling the mixed solution.
More Like This:
Inventors:
MIMORI SHIGETAKA (JP)
MITSUMATA RYOTARO (JP)
SAKAI KENTARO (JP)
KUROSAWA DAISUKE (JP)
MITSUMATA RYOTARO (JP)
SAKAI KENTARO (JP)
KUROSAWA DAISUKE (JP)
Application Number:
PCT/JP2013/078470
Publication Date:
April 24, 2014
Filing Date:
October 21, 2013
Export Citation:
Assignee:
DENKA SEIKEN KK (JP)
International Classes:
C12N15/09; C12Q1/68
Domestic Patent References:
| WO1997022700A2 | 1997-06-26 |
Other References:
ITURRIZA-GOMARA M. ET AL.: "Comparison of specific and random priming in the reverse transcriptase polymerase chain reaction for genotyping group A rotaviruses", J. VIROL. METHODS, vol. 78, no. 1-2, 1999, pages 93 - 103
GUO K. ET AL.: "Development of TaqMan real-time RT-PCR for detection of avian reoviruses", J. VIROL. METHODS, vol. 177, no. L, 2011, pages 75 - 79
ATTOUI H. ET AL.: "Strategies for the sequence determination of viral dsRNA genomes", J. VIROL. METHODS, vol. 89, no. 1-2, 2000, pages 147 - 158
ESCAFFRE O. ET AL.: "Development and validation of four Real-Time quantitative RT-PCRs specific for the positive or negative strands of a bisegmented dsRNA viral genome, J", VIROL. METHODS, vol. 170, no. 1-2, 2010, pages 1 - 8
LAMBDEN PR. ET AL.: "Cloning of viral double- stranded RNA genomes by single primer amplification", METHODS IN MOLECULAR GENETICS, vol. 7, 1995, pages 359 - 372
GUO K. ET AL.: "Development of TaqMan real-time RT-PCR for detection of avian reoviruses", J. VIROL. METHODS, vol. 177, no. L, 2011, pages 75 - 79
ATTOUI H. ET AL.: "Strategies for the sequence determination of viral dsRNA genomes", J. VIROL. METHODS, vol. 89, no. 1-2, 2000, pages 147 - 158
ESCAFFRE O. ET AL.: "Development and validation of four Real-Time quantitative RT-PCRs specific for the positive or negative strands of a bisegmented dsRNA viral genome, J", VIROL. METHODS, vol. 170, no. 1-2, 2010, pages 1 - 8
LAMBDEN PR. ET AL.: "Cloning of viral double- stranded RNA genomes by single primer amplification", METHODS IN MOLECULAR GENETICS, vol. 7, 1995, pages 359 - 372
Attorney, Agent or Firm:
THE PATENT CORPORATE BODY ARUGA PATENT OFFICE (JP)
Patent business corporation Alga patent firm (JP)
Patent business corporation Alga patent firm (JP)
Download PDF: