Co-modified nucleic acid segment of locked nucleic acid and minor groove conjugation

A technology of nucleic acid fragments and co-modification, which is applied in the direction of DNA/RNA fragments, sugar derivatives, biochemical equipment and methods, etc. It can solve the problems of limited annealing temperature and inability to fully meet the design requirements, and achieve the effect of enhanced recognition ability

Active Publication Date: 2010-03-03
SHANGHAI ZJ BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these two methods cannot fully meet the design requirements because of their limited increase in annealing temperature.

Method used

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  • Co-modified nucleic acid segment of locked nucleic acid and minor groove conjugation
  • Co-modified nucleic acid segment of locked nucleic acid and minor groove conjugation
  • Co-modified nucleic acid segment of locked nucleic acid and minor groove conjugation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1: Take Taqman probe as an example

[0058] 1. Design and synthesis of primers and probes

[0059] Primers and probes were designed using the adefovir (ADV) mutation of hepatitis B virus as a detection template.

[0060] Design principles: 1) The primers are designed to amplify a nucleotide sequence containing adefovir (ADV) mutations according to the primer design principles.

[0061]2) The probe is designed with the mutant sequence as the target sequence.

[0062] The goal of the design: in hepatitis B positive samples, samples without adefovir (ADV) mutation cannot release fluorescent signals because the probe cannot bind to it; on the contrary, the mutant samples combine with the target nucleotide fragments to generate fluorescent signals .

[0063] Primer: Forward primer: ADV-236-1: 5'-TGGGCCTCAGTCCGTTTC-3' (SEQ ID NO: 1)

[0064] Reverse primer.: ADV-236-2: 5'-CCAATTACATATCCCATGAAGTTAAG-3' (SEQ ID NO: 2)

[0065] 1) common Taqman probe, 2) MGB prob...

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Abstract

The invention relates to a marking method of oligonucleotide and discloses a co-modified nucleic acid segment of a locked nucleic acid (LNA) and a minor groove binding agent (MGB), namely, both the LNA and the MGB are marked on the same oligonucleotide chain. The invention enables that the capacity of identifying nucleotide sequence mutation and SNP of the oligonucleotide is enhanced by applying the combined modification of the MGB and the LNA.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an oligonucleotide labeling method. Background technique [0002] In order to ensure the binding efficiency of the probe or primer to the template during the PCR amplification process, both the probe and the primer have a certain length limitation. Generally, the length of primers designed is 18-25 bases, while the length of TaqMman probes is generally 20-40 bases. However, in the actual process, in the detection of some mutation sites, SNPs and highly mutated viruses, general probes and primers often have unsatisfactory detection results, poor sensitivity and specificity. [0003] The full name of MGB is minor groove binder, which is a chemical group derived from certain antibiotic molecules. It can be embedded in the minor groove in the DNA double helix structure to form a non-covalent bond. Generally, it can anneal the oligonucleotide Increasing the temperature by more than 10 d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/11C12Q1/68
CPCC07H21/00C12Q1/6818
Inventor 邵俊斌沈步超倪卫琴赵红喜朱勤玮张彬彬马丽丽舒峰朱旭平
Owner SHANGHAI ZJ BIO TECH
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