Improved method for simultaneous identification of differentially expressed mRNAs and measurement of relative concentrations
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 comprise...
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Main Authors | , |
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Format | Patent |
Language | English |
Published |
16.02.2006
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Subjects | |
Online Access | Get full text |
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Summary: | 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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Bibliography: | Application Number: AU19990048497 |