Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Multiple detection method for DNA polymorphism of genome and special probe thereof

A polymorphism and genome technology, applied in the field of genetic engineering, can solve the problem of low throughput

Inactive Publication Date: 2013-10-23
INST OF BOTANY CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method for detecting insertion / deletion markers is mainly to design a pair of PCR primers for an insertion / deletion site, and through PCR amplification, gel or capillary electrophoresis detection, generally only one insertion / deletion marker can be detected at a time, and the throughput is very high. Low

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Multiple detection method for DNA polymorphism of genome and special probe thereof
  • Multiple detection method for DNA polymorphism of genome and special probe thereof
  • Multiple detection method for DNA polymorphism of genome and special probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Embodiment 1, the button hand probe of single nucleotide polymorphism (SNP)

[0111] 1. Detection of two SNPs in the genome of Aegilops tauschii

[0112] 1. Design of button probes PdPCJ819266A, PdPCJ819266G, PdPBJ262615T, PdPBJ262615C

[0113] The genome of Aegilops tauschii is the donor of the D genome of common wheat (Triticum aestivum). PdPCJ819266 and PdPBJ262615 are two SNPs of Aegilops tauschii (Aegilops tauschii), in which AA and TT are genotypes of Aegilops tauschii variety 2280, and GG and CC are genotypes of Aegilops tauschii variety 2282, that is, Aegilops tauschii 2280 and Aegilops tauschii The 62nd nucleotide in the sequence 5 of the 2282 variety has a polymorphism, which are AA and GG respectively; the 175th nucleotide in the sequence 6 of the 2280 and 2282 varieties of A. tachyphylla has a polymorphism, which are respectively TT and CC.

[0114] Among them, the button-hand probe PdPCJ819266A specifically recognizes allele A with a length of 147 nt, th...

Embodiment 2

[0161] Embodiment 2, insertion / deletion polymorphism (InDel) labeled button hand probe

[0162] 1. Detection of two insertion / deletion (InDel) markers in the barley genome

[0163] 1. Design of button probes PdPInDel1-1, PdPInDel1-2, PdPInDel2-1, PdPInDel2-2

[0164] PdPInDel1 and PdPInDel2 are two InDel markers on the fifth chromosome (5H) of barley (Hordeum vulgare L.), wherein InDel1-1 and InDel2-1 are specific genotypes of barley variety Vada, and InDel1-2 and InDel2-2 are Genotypes specific to barley cultivar L94. The button-hand probe PdPInDel1-1 specifically recognizes the InDel1-1 genotype, and its length is 160 nt. The button-hand probe PdPInDel1-2 specifically recognizes the InDel1-2 genotype, and its length is 190 nt. The genotype is 318nt in length, and the clasp probe PdPInDel2-2 specifically recognizes the InDel2-2 genotype with a length of 342nt.

[0165] The 5' ends of the four hand probes were all phosphorylated. See Table 4 for sequence design and structur...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

Provided are a method for multiplex detection of genomic DNA polymorphism and probes dedicated thereto. Also disclosed is a single-stranded DNA molecule comprising successively, from the 5' end to the 3' end, a left-side oligonucleotide fragment complementary to the 5' end of the DNA to be detected, a main fragment and a right-side oligonucleotide fragment complementary to the 3' end of the DNA to be detected. Through ligase reaction, different labels and different allelic sites are distinguished by the single-stranged DNA molecule. Hand-clasping probes of different lengths are successfully ligated by PCR amplification. Through gel or capillary electrophoresis separation and detection systems, simultaneous detection of the polymorphism of a multiple of deoxyribonucleic acids is achieved. High sensitivity and accuracy are attained, and the requirements for high-throughput and low cost are satisfied at the same time.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for multiple detection of genomic DNA polymorphism and a special probe thereof. Background technique [0002] Deoxyribonucleic acid (DNA) polymorphism refers to the difference in nucleotide arrangement in chromosomal DNA alleles, that is, there are two or more genotypes in the alleles (or fragments) in the DNA region, which can be divided into mononuclear Nucleotide polymorphism (single nucleotide polymorphism, SNP) and insertion / deletion polymorphism (insertion / deletion, In / Del). [0003] (1) Single Nucleotide Polymorphism (SNP) [0004] Single nucleotide polymorphism (SNP) refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the chromosome genome level, in which at least one allele occurs in no less than 1% of the population. The polymorphism shown by SNP only involves the variation of a single base, which can be caused by the t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6858C12N15/11
Inventor 漆小泉曹晓花
Owner INST OF BOTANY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com