METHOD OF INCREASING SPECIFICITY OF REVERSE TRANSCRIPTION

摘要

FIELD: chemistry.;SUBSTANCE: method involves preparation of a sample, presumably containing at least one type of RNA (target) to undergo reverse transcription and at least one type of RNA different from the target; treatment thereof with polynucleotide phosphorylase Thermus thermophilus (Tth PNPhase) at temperature 60-75°C and micromolar concentration of magnesium, carried out after annealing the analysed sample with a protective oligonucleotide lying on the 3'-side of the section to undergo reverse transcription, and forming a hybrid with the target which is stable in incubation conditions; and a reverse transcription reaction which is carried out with the reaction mixture obtained at the preliminary treatment step, without further purification thereof.;EFFECT: high specificity of reverse transcription.;6 cl, 7 dwg, 11 ex