Title:
【発明の名称】弁別的に発現されたmRNAの同時同定および相対濃度測定のための改良方法
Document Type and Number:
Japanese Patent JP2002519011
Kind Code:
A
Abstract:
An improved method for the simultaneous sequence-specific identification of mRNAs in a mRNA population allows the visualization of nearly every mRNA expressed by a tissue as a distinct band on a gel whose intensity corresponds roughly to the concentration of the mRNA. In general, the method comprises the formation of cDNA using anchor primers to fix a 3'-endpoint, producing cloned inserts from the cDNA in a vector containing a bacteriophage-specific promoter for subsequent RNA synthesis, generating linearized fragments of the cloned inserts, preparing cRNA, transcribing cDNA from the cRNA using a set of 5'-RT primers, and performing PCR using a 3'-PCR primer whose sequence is derived from the vector and a set of 5'-PCR primers that is derived from the 5'-RT primers used for transcription of cDNA from cRNA. The method can identify changes in expression of mRNA associated with the administration of drugs or with physiological or pathological conditions.
Inventors:
Sato Cliff Jay Gregaud
Hasel Karl Ve
Hasel Karl Ve
Application Number:
JP2000556998A
Publication Date:
July 02, 2002
Filing Date:
June 30, 1999
Export Citation:
Assignee:
THE SCRIPPS RESEARCH INSTITUTE
International Classes:
C12N15/10; C12P19/34; C12N15/09; C12Q1/68; C12Q1/6809; C12Q1/6813; C12Q1/6853; C12Q1/70; G01N33/48; (IPC1-7): C12N15/09; C12Q1/68
Attorney, Agent or Firm:
Minoru Nakamura (9 outside)