| 摘要 |
FIELD: chemistry.SUBSTANCE: invention relates to biotechnology, specifically to a reagent for preventing at least one false start in polymerase chain reaction (PCR) amplification, a method of preventing at least one false start in PCR amplification and sets which include said reagent. The reagent is used in PCR which is carried out in the presence of 1.25 units of thermally stable DNA polymerase per 25 mcl of the reaction mixture with concentration of not more than 650 nM. The reagent is a non-extending DNA polymerase oligonucleotide, having a barrel-loop structure and is not a hybridisation probe for said amplified DNA product. The barrel has a length greater than six nucleotides, is stabilised at the end far from the loop by non-fluorescent substances selected from: fluorescence extinguisher and a nucleotide analogue which facilitates a tighter connection between oligonucleotides in the barrel than a DNA-DNA hybrid. The melting point of the barrel (Tm) is below 94°C. The loop is an oligonucleotide and/or carbon linker.EFFECT: invention enables to prevent false start (unintentional hybridisation) in amplifications and analyses using PCR.20 cl, 25 dwg, 3 tbl, 14 ex |
