TEMPLATE-DIRECTED ENZYMATIC DNA SYNTHESIS USING PHOSPHORYL GUANIDINE OLIGONUCLEOTIDES

摘要

The invention relates to the field of molecular biology, biotechnology and molecular diagnostics. The invention can be used in the development and optimisation of PCR and RT-PCR systems used for detecting nucleic acids, inter alia, for diagnostics of genetic, viral and other diseases. The essence of the proposed method is comprised in that neutral oligonucleotide derivatives, more specifically phosphoryl guanidines, comprising one or more phosphate groups in which a guanidine or a substituted guanidine residue is inserted at the phosphorus atom, are used as primers in the polymerase chain reaction (PCR) and PCR combined with reverse transcription (RT-PCR). The invention makes it possible to obtain more reliable, specific and selective results when conducting PCR, inter alia, to increase PCR sensitivity by reducing formation of DNA amplification by-products and/or to control the PCR-product yield, including by deliberate suppression, through the use of various combinations of positions and numbers of the modified phosphate groups in the formulation of the oligonucleotide primers.