EP 1127159 A1 20010829 - METHOD FOR INDEXING AND DETERMINING THE RELATIVE CONCENTRATION OF EXPRESSED MESSENGER RNAs
Title (en)
METHOD FOR INDEXING AND DETERMINING THE RELATIVE CONCENTRATION OF EXPRESSED MESSENGER RNAs
Title (de)
VERFAHREN ZUR INDEXIERUNG UND ZUR BESTIMMUNG DER RELATIVEN KONZENTRATION VON EXPRIMIERTEN MRNAS
Title (fr)
PROCEDE PERMETTANT D'INDEXER ET DE DETERMINER LA CONCENTRATION RELATIVE D'ARN MESSAGERS EXPRIMES
Publication
Application
Priority
- US 9923655 W 19991014
- US 18686998 A 19981104
Abstract (en)
[origin: WO0026406A1] 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 and performing two sequence specific PCR amplifications of the cDNA. In preferred embodiments, the method comprises comparing the length and at least part of the nucleotide sequence of the PCR products to expected values determined from a database of nucleotide sequences. The method can identify changes in expression of mRNA associated with the administration of drugs or with physiological or pathological conditions. Also provided are vectors and primers useful for the practice of the improved method.
IPC 1-7
IPC 8 full level
C12N 15/09 (2006.01); C12Q 1/68 (2006.01); C12Q 1/6809 (2018.01); G01N 33/15 (2006.01); G01N 33/50 (2006.01)
CPC (source: EP KR)
C12Q 1/68 (2013.01 - KR); C12Q 1/6809 (2013.01 - EP)
Citation (search report)
See references of WO 0026406A1
Designated contracting state (EPC)
AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE
DOCDB simple family (publication)
WO 0026406 A1 20000511; AU 1108900 A 20000522; CA 2350168 A1 20000511; CN 1331755 A 20020116; EA 200100490 A1 20011022; EP 1127159 A1 20010829; IL 142965 A0 20020421; JP 2002528135 A 20020903; KR 20010092721 A 20011026; MX PA01004550 A 20020918; NO 20012203 D0 20010503; NO 20012203 L 20010702
DOCDB simple family (application)
US 9923655 W 19991014; AU 1108900 A 19991014; CA 2350168 A 19991014; CN 99814965 A 19991014; EA 200100490 A 19991014; EP 99954838 A 19991014; IL 14296599 A 19991014; JP 2000579778 A 19991014; KR 20017005872 A 20010504; MX PA01004550 A 19991014; NO 20012203 A 20010503