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A kind of method and application of rapid detection Saccharum spsb gene polymorphism

A gene polymorphism, SPSB2177P2 technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems such as the intron region has not been cloned, and the intron region polymorphism analysis has not been reported. , to achieve the effect of overcoming high cost, high accuracy and strong primer specificity

Inactive Publication Date: 2016-04-13
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are many studies on the cloning and function of the sugarcane SPSB gene at home and abroad, mainly from the perspective of mRNA, but the intron region of the gene has not been completely cloned, and the polymorphism of the intron region Analysis has not been reported

Method used

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  • A kind of method and application of rapid detection Saccharum spsb gene polymorphism
  • A kind of method and application of rapid detection Saccharum spsb gene polymorphism
  • A kind of method and application of rapid detection Saccharum spsb gene polymorphism

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Embodiment 1

[0047] (1) Cloning and polymorphism detection of partial DNA sequences of SPSB genes in different species of Saccharum.

[0048] 1. Extraction of sample DNA

[0049] In the present invention, sugarcane large-stem wild species 51NG3, Indian species Nagori, sugarcane variety ROC22, tropical species Badila, Chinese species glossy bamboo cane and sugarcane thin-stem raw species Yincut No. 1 are used as detection objects, and are extracted with the EasyPurePlantGenomicDNAKit provided by Quanshijin Company Genomic DNA was finally dissolved in sterilized ultrapure water and stored at 4°C after detection. The OD value of the DNA samples was measured with a UV photometer. Calculate the DNA content and the ratio of OD260 / OD280. If the ratio of OD260 / OD280 is less than 1.6, it means that the sample contains more protein or phenol, and purification should be performed; if the ratio is greater than 1.8, RNA purification should be considered. After the DNA detection is completed, a certai...

Embodiment 2

[0072] (1) Cloning of partial DNA sequences of SPSB genes of different species of Saccharum and detection of polymorphisms

[0073] 1. Extraction of sample DNA

[0074] In the present invention, the wild sugarcane species 51NG3, the Indian species Nagori, the sugarcane variety ROC22, the tropical species Badila, and the Chinese species glossy bamboo cane are used as detection objects, and the Genomic DNA is extracted with the EasyPurePlantGenomicDNAKit provided by Quanshijin Company. Dissolve in pure water and store at 4°C after detection. The OD value of the DNA samples was measured with a UV photometer. Calculate the DNA content and the ratio of OD260 / OD280. If the ratio of OD260 / OD280 is less than 1.6, it means that the sample contains more protein or phenol, and purification should be performed; if the ratio is greater than 1.8, RNA purification should be considered. After the DNA detection is completed, a certain amount is taken out and diluted to 40ng / μL for use as a P...

Embodiment 3

[0098] (1) Cloning and polymorphism detection of the partial DNA sequence of the SPSB gene of the wild sugarcane thin-stalked species Yinque 1.

[0099] 1. Extraction of sample DNA

[0100] In the present invention, the sugarcane thin-stem seed Yinke No. 1 is used as the detection object, and the Genomic DNA is extracted with the EasyPurePlantGenomicDNAKit provided by Quanshijin Company, and finally dissolved in sterilized ultrapure water, and stored at 4°C after detection. The OD value of the DNA samples was measured with a UV photometer. Calculate the DNA content and the ratio of OD260 / OD280. If the ratio of OD260 / OD280 is less than 1.6, it means that the sample contains more protein or phenol, and purification should be performed; if the ratio is greater than 1.8, RNA purification should be considered. After the DNA detection is completed, a certain amount is taken out and diluted to 40ng / μL for use as a PCR template.

[0101] 2. Primer design

[0102] Obtain the publish...

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Abstract

The invention discloses a method for rapidly detecting the polymorphism of a saccharum sucrose phosphate synthase B (SPSB) gene, belonging to the field of molecular breeding of plants. The method comprises the steps of designing a pair of primers to carry out PCR amplification by taking the genome DNA of saccharum robustum B.51NG3, saccharum barberi J.Nagori, saccharum species ROC22, saccharum.officinarum L.Badila, saccharum sinense R. lustrous bamboocane or saccharum spontaneum L.yinge No.1 as a template; carrying out agarose gel detection to name the SPSB genotype of a fragment with the size of 287bp as type C1 and name the SPSB genotype of a fragment with the size of 331bp as type C2; and identifying the polymorphism of the saccharum SPSB gene according to different sizes of the fragments. The invention provides a DNA level identifying method for saccharum sugar trait marker-assisted selection, and the DNA level identifying method is beneficial to tracking and utilization of different haplotypes of SPSB genes in filial generations of saccharum and has important significance for rapidly establishing saccharum germplasm with excellent heredity.

Description

technical field [0001] The invention belongs to the technical field of plant molecular breeding, and in particular relates to a method for detecting the polymorphism of sugarcane SPSB gene (JN584485), and its application in marker-assisted selective breeding of sugarcane sugar traits to rapidly establish genetically excellent sugarcane germplasm. Background technique [0002] Saccharum (Saccharum) includes tropical species (Saccharum.officinarum L.), Chinese species (SaccharumsinenseR.), Indian species (SaccharumbarberiJ.), sugarcane large stem wild species (Saccharum robustumB.) and sugarcane thin stem wild species (SaccharumspontaneumL.), is Sugarcane resources commonly used in sugarcane hybrid breeding have complex and variable chromosomes, and most of them are allopolyploid plants, with multiple sets of chromosomes in the nucleus. Therefore, the traits of hybrid progeny are seriously separated, and the progeny of a hybrid panicle has hundreds of progeny types, and the tr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘洪博刘新龙范源洪蔡青陆鑫毛钧徐超华李旭娟苏火生马丽林秀琴
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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