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 optimization of PCR and RT-PCR systems used to detect nucleic acids, including the diagnosis of genetic, viral, and other diseases. The essence of the proposed method is that neutral derivatives of oligonucleotides, namely phosphoryl guanidines containing one or more phosphate groups in which guanidine or substituted guanidine residue is introduced on the phosphorus atom, are used as primers for the template-based amplification, including polymerase chain reaction (PCR) and PCR combined with reverse transcription (RT-PCR). The invention allows to obtain more reliable, specific and selective results in the process of PCR, in particular, to increase the sensitivity of PCR by reducing the yield of by-products of DNA amplification and/or to control the yield of the PCR product, including intentionally suppressing, by using different combinations of the location and number of modified phosphate groups in the oligonucleotide primers.