Detection method for single nucleotide polymorphism

A single nucleotide polymorphism and detection method technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high cost of detection equipment, isotope pollution, etc.
CN103103283AInactive Publication Date: 2013-05-15CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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Patent Information

Authority / Receiving Office
CN Β· China
Patent Type
Applications(China)
Current Assignee / Owner
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
Publication Date
2013-05-15
Estimated Expiration
Not applicable Β· inactive patent

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Abstract

The invention belongs to the technical field of molecular biology, relates to analysis and detection for single nucleotide polymorphism, and in particular relates to a universal detection method. The detection method comprises the steps of amplifying target DNA (deoxyribonucleic acid) by gap-ligase chain reaction, introducing a DNAzyme sequence into a ligation product, degrading a phosphorylated probe which does not participate in the ligation reaction by Lambda excision enzyme, and releasing DNAzyme by excision enzyme III. The detection method is characterized in that a DNAzyme probe is combined with a Gap-LCR (ligase chain reaction) probe, and is characterized by comprising the following steps of: adding a section of DNAzyme sequence into a phosphorylated probe to be connected in an LCR system, adding an Anti-G sequence into a system (in complementary pairing with a DNAzyme sequence part in the probe) after the Gap-LCR amplification reaction stops, degrading a 5-end phosphorylated DNA probe by the Lambda excision enzyme, and releasing the DNAzyme in the ligation product by the excision enzyme III, thereby qualitatively and quantitatively detecting the SNP (single nucleotide polymorphism) due to the characteristics of the DNAzyme. The method is simple, convenient, practical and economical, and is widely applicable to the detection of various single nucleotide polymorphisms.
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Description

technical field

[0001] The invention belongs to the technical field of molecular biology and relates to the analysis and detection of single nucleotide polymorphisms, in particular to a universal method for amplifying target DNA by gap-ligase chain reaction, introducing DNAzyme sequences into ligation products, and then A detection method that uses Lambda exonuclease to degrade phosphorylated probes that do not participate in the ligation reaction, and exonuclease III to release DNAzyme. Background technique

[0002] Single nucleotide polymorphism (single nucleotide polymorphism, SNP), mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genomic level, which is a specific nucleotide position in genomic DNA. Changes such as conversion, transversion, insertion or deletion occur, and the frequency of any allele in the population is not less than 1%. It is the most common type of human heritable variation, accounting for more than...

Claims

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