Cucumber SNP marker and detection methods thereof

A detection method and technology for cucumber, which are applied in the fields of biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, can solve the problems of difficulty, high cost, and restriction on the application of SNP markers in the identification of SNP marker sites. , to achieve the effect of shortening development and verification time and reducing inspection costs

Inactive Publication Date: 2009-12-02
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

Traditional identification and detection methods are time-consuming, labor-intensive and costly in this large-throughput analysis
This also restricts the application of SNP markers in plant molecular genetics research to a certain extent.
At the same time, for some plant materials that are not rich in genome sequence information (such as the cucumber material used in the present invention), the identification of SNP marker sites itself also has certain difficulties.

Method used

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  • Cucumber SNP marker and detection methods thereof
  • Cucumber SNP marker and detection methods thereof
  • Cucumber SNP marker and detection methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A method for rapid identification of SNP loci

[0029] Step 1: Sequencing the target DNA interval on the cucumber parent materials to determine the candidate SNP sites; then the F 1 Sequencing of the target DNA interval in a single generation

[0030] In the present invention, the PCR marker is used to scan and screen the cucumber genome BAC library, and then obtain the related genome sequence linked with the M gene. All PCR reaction systems are: BAC library plasmid 20ng, bidirectional primers 0.2μmol / L, 200μmol / L dNTPs, 2mmol / L MgCl 2 , 1×Taq buffer, 0.5UT Taq DNA polymerase, the total reaction system is 10 μL, and Taq DNA polymerase was purchased from Promega Company. The amplification program was: 94°C for 3min; 40cycles, 94°C for 20s, 65°C for 30s, 72°C for 30s; 72°C for 5min, and the results were displayed by 1.5% agarose gel electrophoresis.

[0031] The present invention first uses the marker S_ME8SA7 to screen the expanded population, obtains two exchanged in...

Embodiment 2

[0037] Construction of genetically isolated populations and identification of exchanged individual plants

[0038] Step one, F 2 Segregation Population Construction

[0039] European greenhouse type inbred line H34 (male parent), hermaphrodite flower variety, the whole plant bears hermaphrodite flowers under normal growth conditions, the plant grows faster, mature fruit is spherical, dark green; South China type inbred line S52 (female parent) This), a unisexual flowering variety, in the early stage of plant growth (lower node position) male flowers are born, then male flowers and female flowers are distributed alternately, in the late growth period (higher node position) female flowers are completely born, the plant growth rate is slow, and the mature fruit is long. shape, white. F 1 Progeny, unisexual flowering plant (strong female plant), long strip-shaped fruit, dark green. Inbred lines S52, H34, F 1 Judgment criteria for offspring and unisexual flowering plants / bisex...

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Abstract

The invention relates to a cucumber SNP marker and detection methods thereof in the technical field of plant gene engineering. A method for identifying cucumber SNP locus comprises the following steps: selecting homozygous cucumber parents, and performing sequencing to determine candidate SNP locus; hybridizing to obtain F1 generation single plants, and performing sequencing; and detecting a peak shape chart of a F1 generation sequencing result to determine candidate locus with heterozygous peaks. A method for determining SNP marked polymorphic single plants in F2 generation separation group of cucumber comprises the following steps: selecting homozygous cucumber parents, and obtaining the F2 generation separation group; selecting recessive phenotype single plants in the F2 generation, mixing DNAs of the single plants in equal quantity, and construct a recessive gene pool; performing amplification and sequencing by an SNP primer, and detecting a peak shape chart of a sequencing result; and respectively performing amplification and sequencing on single plants in the recessive gene pool by the SNP primer, and detecting a peak shape chart of a sequencing result. The cucumber SNP marker has a sequence shown as SEQ ID NO:1. The invention shortens the development and testing time of the SNP marker, and reduces the detection cost of the marker.

Description

technical field [0001] The invention relates to a marker in the technical field of genetic engineering and a detection method thereof, in particular to a cucumber SNP (single nucleotide polymorphism) marker and a detection method thereof. Background technique [0002] A molecular marker in a broad sense refers to a specific DNA sequence, RNA sequence or protein that can be inherited and detected. Molecular markers in a narrow sense only refer to DNA markers, and this definition has been widely adopted around the world. Molecular markers are genetic markers that reflect genetic variation at the DNA molecular level. It has two basic characteristics, namely heritability and recognizability. The emergence and wide application of DNA molecular markers have greatly promoted the development of gene mapping and genome mapping, making it an important field of genetics and even the entire biological research. Since geneticist Botstein first proposed the idea of ​​DNA restriction fr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 蔡润李征潘俊松何欢乐曹娴
Owner SHANGHAI JIAO TONG UNIV
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