FIELD: medicine; biotechnology.
SUBSTANCE: group of inventions refers to molecular biology, organic synthesis, enzymology, biotechnology and medicine. Disclosed is a method for enzymatic production of modified single-stranded DNA to create reagents capable of specific binding to hydrophobic sections of high-molecular organic compounds. Method is based on polymerase chain reaction, either transcription, or reverse transcription, or primer extension reaction using modified deoxyuridin triphosphates, unmodified DNA sequences with a given sequence, highly specific flanking primers, where 3'-primer is biotinylated at 5'-end, KOD XL or Deep Vent polymerase. Modified DNA strands are separated from unmodified DNA chains using magnetic particles with covalently cross-linked streptavidin molecules to their surface, or by using λ-exonuclease.
EFFECT: obtained single-stranded DNA with modifications are able to interact with hydrophobic sections of high-molecular target compounds and are intended for development of DNA probes, aptamers and somamers used in molecular biological studies and in medical diagnostics.
12 cl, 5 dwg, 2 tbl, 21 ex
Title | Year | Author | Number |
---|---|---|---|
METHOD FOR DETERMINING SUBSTRATE EFFICIENCY OF DEOXYURIDINE TRIPHOSPHATES DERIVATIVES FOR DNA POLYMERASES BY PRIMER COMPLETION REACTION | 2019 |
|
RU2731739C1 |
DEOXYURIDINE TRIPHOSPHATES BOUND TO CYANINE DYES BY SULFAMIDOALKYL LINKERS, FOR USE IN PCR | 2017 |
|
RU2667070C1 |
FLUORESCENTLY-LABELED DEOXYURIDINE TRIPHOSPHATES | 2016 |
|
RU2637310C1 |
METHOD FOR DETERMINING SUBSTRATE COMPATIBILITY OF MODIFIED DEOXYURIDINE TRIPHOSPHATE AND DNA POLYMERASE BY COMBINING QUANTITATIVE PCR AND ELECTROPHORESIS METHODS | 2019 |
|
RU2736969C1 |
DEOXYURIDINE TRIPHOSPHATES MARKED WITH ZWITTERIONIC INDOCYANINE DYES | 2017 |
|
RU2725884C2 |
METHOD FOR EVALUATING THE EFFECT OF MODIFIED DEOXYNUCLEOSIDE TRIPHOSPHATES ON THE OLIGONUCLEOTIDE COMPOSITION OF COMBINATORIAL DNA LIBRARIES FOR SELECTING MODIFIED APTAMERS | 2021 |
|
RU2779058C1 |
THERMAL DISSOCIATION METHOD FOR SELECTION OF DNA-APTAMERS | 2019 |
|
RU2723373C1 |
METHOD OF PRODUCING MODIFIED COMBINATORIAL DNA LIBRARIES BY SOLID-PHASE PRIMER EXTENSION REACTION ON PET MICROPARTICLES | 2018 |
|
RU2734941C2 |
METHOD FOR CREATION AND SELECTION OF THE LIBRARY OF MODIFIED APTAMERS | 2016 |
|
RU2652952C1 |
METHOD FOR DETERMINATION OF NUCLEIC ACID MOLECULE SEQUENCE CHARACTERISTICS VIA DETECTION AND CHARACTERIZATION OF SAID SEQUENCE VARIATIONS | 1998 |
|
RU2264468C2 |
Authors
Dates
2019-09-05—Published
2017-11-30—Filed