Title:
微量mRNAの増幅方法およびその利用
Document Type and Number:
Japanese Patent JP5078038
Kind Code:
B2
Abstract:
In order to provide a method for amplifying a trace amount of mRNA which can amplify a short mRNA and a long mRNA at a same efficiency level, regardless of how long a base sequence is, and to provide a method for use of such a method, a method of the present invention for amplifying a trace amount of mRNA includes the steps of: (i) adding a dummy RNA to a solution containing the trace amount of mRNA, so as to prepare a mixed solution; (ii) synthesizing an anti-sense DNA by reverse transcription, which uses the mixed solution as a template; (iii) synthesizing a sense DNA which is complementary to the anti-sense DNA thus synthesized, so as to generate a double strand DNA made of the sense DNA and the anti-sense DNA; (iv) ligating an RNA polymerase promoter sequence to the double strand DNA thus generated, on a sense DNA 5' end side of the double strand DNA, so as to prepare a double strand DNA for amplification; and (v) amplifying, by using RNA polymerase, an RNA from the double strand DNA for amplification.
Inventors:
Hiroshi Nojima
Tokihiro Togishi
Okuzaki Daisuke
Tokihiro Togishi
Okuzaki Daisuke
Application Number:
JP2008534286A
Publication Date:
November 21, 2012
Filing Date:
August 29, 2007
Export Citation:
Assignee:
National University Corporation Osaka University
International Classes:
C12N15/09; C12Q1/68
Domestic Patent References:
| JP2002238575A | 2002-08-27 |
Attorney, Agent or Firm:
Kenzo Hara International Patent Office