METHOD FOR POLYMERASE ACTIVITY DETERMINATION

• Main Authors: HAGSTROEM, ASA, MARGARETA, NEUMUELLER, THORD, MAGNUS, GRONOWITZ, JAN-SIMON, KAELLANDER, CLAS, FREDRIK, RUNESSON, KARLSTROEM, ROLF, ANDERS, LANGSTROEM-PERSSON, ULLA, M, WELTMAN, EVA, HELENA
• Format: Patent
• Language: English; French; German
• Published: 21.09.1994
• Edition: 5
• Subjects: ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING; BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS,e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES,DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; CONVEYING; DERIVATIVES THEREOF; ENZYMOLOGY; HANDLING THIN OR FILAMENTARY MATERIAL; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MAKING ARTICLES OF PAPER, CARDBOARD OR MATERIAL WORKED IN A MANNERANALOGOUS TO PAPER; MAKING CONTAINERS OF PAPER, CARDBOARD OR MATERIAL WORKED IN AMANNER ANALOGOUS TO PAPER; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC CHEMISTRY; PACKAGES; PACKAGING ELEMENTS; PACKING; PERFORMING OPERATIONS; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDEDFOR; SHAPING OR JOINING OF PLASTICS; SPIRITS; STORING; SUGARS; TESTING; TRANSPORTING; VINEGAR; WINE; WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE, IN GENERAL; WORKING PAPER, CARDBOARD OR MATERIAL WORKED IN A MANNER ANALOGOUSTO PAPER

The invention relates to a method of quantitatively determining nucleic acid polymerase activity in a sample, comprising incubation of the sample with a natural or completely or partially synthetic polynucleotide template and a reagent solution containing the necessary substrates including at least one radiolabeled nucleoside triphosphate complementary to the template, separation of the template from the substrate, and measurement of the radioactivity incorporated into the template, which radioactivity is substantially proportional to the polymerase activity in the sample, and wherein, when a primer is required, the latter is either hybridized to the template from the start or added during the determination. According to the invention the template is immobilized to a carrier, said nucleoside triphosphate is labeled with a gamma-radiating isotope, and the measurement of the incorporated radioactivity is performed directly on the immobilized template without release thereof from the carrier. The invention also relates to a kit for determining of nucleic acid polymerase activity, comprising a carrier-bound template and a substrate labeled with a gamma-radiating isotope.