Preparation method and application of oligonucleotide with closed dideoxyribonucleoside

A nucleotide and nucleic acid technology, applied in biochemical equipment and methods, microbiological measurement/testing, etc., can solve problems such as poor sensitivity and accuracy, time-consuming and cumbersome operations, and complicated operations
CN109735607AInactive Publication Date: 2019-05-10INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
Publication Date
2019-05-10
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a preparation method and application of an oligonucleotide with a closed dideoxyribonucleoside. The method specifically comprises the steps that a terminal transferase (TdT) isfirstly used for catalyzing deoxyribonucleotides (dNTPs) or dideoxynucleotides (ddNTPs) in combination with the characteristics of a 3'hydroxyl terminal of a DNA molecule, so that a primer is mixed with any one of four dideoxynucleotides (ddATP, ddTTP, ddCTP or ddGTP), the TdT can add the dideoxynucleotides to the 3'hydroxyl terminal of the primer, and the obtained primer modified by the ddNTP cannot be catalytically extended by a DNA polymerase. The prepared oligonucleotide (primer) with the closed dideoxyribonucleoside can be used in combination with amplification methods such as allele PCR,proof-reading PCR for detecting mutation.
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Description

technical field

[0001] The invention belongs to the technical field of nucleic acid detection, and more specifically relates to a preparation method and application of a dideoxynucleoside-blocked oligonucleotide. Background technique

[0002] Point mutation, also known as single base substitution, refers to a mutation caused by a single base change, which can be divided into two types: transition and transversion. A transition refers to a substitution between a purine and a purine or a pyrimidine to a pyrimidine; a transversion refers to a substitution between a purine and a pyrimidine. Single-base mutations in DNA molecules widely exist in the encoding of genetic information in organisms, and are one of the important causes of genetic diseases. As far as the human genome is concerned, among the sense mutations of many genes that cause human diseases, pathogen subtypes, and drug resistance genes, single-base mutations account for a considerable proportion. The important to...

Claims

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