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PCR-Reaktionsgemisch fuer Fluoreszenz-basierende Genexpressions- und Genmutationsanalysen

Polymerase chain reaction mixture, useful e.g. for genomic analysis, includes bovine serum albumin, allowing reduction in concentration of polymerase and fluorescent reagents.
: Borlak, J.; Thum, T.

Frontpage ()

DE 2000-10046079 A: 20000915
DE 2001-10147232 A: 20010915
WO 2001-DE3617 A: 20010915
EP 2001-984658 A: 20010915
EP 2001-984658 AW: 20010915
DE 10147232 A1: 20020711
EP 1317570 B1: 20050406
Patent, Elektronische Publikation
Fraunhofer ITEM ()

Der Erfindung liegt die Aufgabe zugrunde, die Selektivitaet und Sensitivitaet Fluoreszenz-basierender Genexpressions- und Genmutationsanalysen zu steigern und Fehldiagnosen und irrtuemliche Befundungen zu verhindern. Das PCR-Reaktionsgemisch fuer Fluoreszenz-basierende Genexpressions- und Genmutationsanalysen ist dadurch gekennzeichnet, dass es neben den ueblichen Reaktionskomponenten zusaetzlich Bovines Serum Albumin enthaelt und eine erhoehte MgCl2-Konzentration aufweist.


WO 200222865 A UPAB: 20020618 NOVELTY - Polymerase chain reaction (PCR) mixture (A) for fluorescent-based gene expression and gene mutation analysis comprising: a Taq polymerase (I); bovine serum albumin (BSA) at final concentration 400-800 mu g/ml; a high content of magnesium chloride, depending on the (I) used, the final concentration is over 3.5 to 5 mM; and a fluorescent material (II). USE - (A) is used: (a) to detect nucleotide polymorphisms, especially in cases of Gilbert's syndrome; (b) to characterize potential pharmaceuticals at the level of gene expression; (c) for pharmacogenomics or toxicogenomics; (d) for molecular diagnosis; (e) for identifying molecular interaction between materials and biological agents at the gene level; and (f) to identify gene interactions and for functional analysis of new genes, including sequencing and cloning. Specific applications are detecting cDNA representing the genes for sarcoplasmic calcium Data, beta -myosin heavy chain, brain or atrial native peptides, or alpha -skeletal actin (all in heart tissue or cultured cardio myocytes); albumin and HF-3c (transcription factor), in cultured hepatocytes; the allele 5 asterisk of N-acetyltransferase-2 in human lymphocyte DNA; or the CYP2J3 gene in explanted rat hearts. ADVANTAGE - Addition of BSA and the increase in magnesium chloride concentration improves selectivity and sensitivity, preventing misdiagnosis and errors. (A) is inexpensive, specifically because the amounts of expensive (II) and Taq (from 2.5 to 0.9 units per assay) can be reduced. The mixture can be used successfully where the nucleic acid concentration is too low for analysis by known methods. 9