Method for simultaneous identification of differentially express

Details Method for simultaneous identification of differentially express Method for simultaneous identification of differentially express

1998-06-30

2000-08-01

Schwartzman, Robert A.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

536 2433, C12Q 168, C07H 2104

Patent

active

060965037

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.

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