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
Title | Year | Author | Number |
---|---|---|---|
METHOD FOR HOMOGENOUS DETECTION OF ONE-STRAND AMPLIFICATION PRODUCT | 2005 |
|
RU2460804C2 |
OLIGONUCLEOTIDE PRIMERS FOR B.mallei GENETIC TYPING BY POLYMERASE CHAIN REACTION | 2012 |
|
RU2478714C1 |
OLIGONUCLEOTIDE PRIMERS FOR GENETIC TYPING B. mallei BY POLYMERASE CHAIN REACTION | 2012 |
|
RU2474618C1 |
OLIGONUCLEOTIDE PRIMERS FOR GENETIC TYPING B. mallei BY POLYMERASE CHAIN REACTION | 2012 |
|
RU2474619C1 |
OLIGONUCLEOTIDE PRIMERS FOR B.mallei GENETIC TYPING BY POLYMERASE CHAIN REACTION | 2012 |
|
RU2478715C1 |
METHOD FOR DETECTING SEQUENCES OF NUCLEOTIDES BY POLYMERASE CHAIN REACTION METHOD IN REAL TIME WITH UNIVERSAL PROBE | 2017 |
|
RU2675378C1 |
COMPOSITIONS FOR THE REACTION OF REVERSE TRANSCRIPTION WITH A HOT START OR FOR A POLYMERASE CHAIN REACTION WITH REVERSE TRANSCRIPTION WITH A HOT START | 2013 |
|
RU2630999C2 |
DIAGNOSTIC SYSTEM FOR DETECTION OF CATTLE LEUKEMIA AND IMMUNE DEFICIENCY PROVIRUS DNA BY MULTIPLEX POLYMERASE CHAIN REACTION | 2015 |
|
RU2615465C2 |
TEST SYSTEM FOR GENOME DETECTION OF ROTAVIRUS AGENT OF TYPE A IN AGRICULTURAL ANIMALS BY MEANS OF MULTIPLEX POLYMERASE CHAIN REACTION WITH FLUORESCENT DETECTION IN REAL TIME | 2018 |
|
RU2694501C1 |
SYNTHETIC OLIGONUCLEOTIDE PRIMERS AND METHOD FOR THEREOF FOR INDICATING IMMUNODEFICIENCY VIRUSES AND LEUKAEMIA OF CATS IN CLINICAL MATERIAL BY MULTIPLEX POLYMERASE CHAIN REACTION | 2015 |
|
RU2586504C1 |
Authors
Dates
2013-10-10—Published
2005-10-17—Filed