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A group of maize ssr molecular markers and their application

A molecular marker, corn technology, applied in the field of crop molecular biology, can solve the problems of equipment and personnel skill requirements, difficult to distinguish clearly, unfavorable popularization, etc., and achieve the effects of high sensitivity, easy judgment, and convenient operation.

Active Publication Date: 2021-02-09
黑龙江省农业科学院农产品质量安全研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low resolution of this technology, it is difficult to clearly distinguish the difference of a few bases between the amplified fragments, and the requirements for equipment and personnel skills are relatively high, which is not conducive to popularization

Method used

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  • A group of maize ssr molecular markers and their application
  • A group of maize ssr molecular markers and their application
  • A group of maize ssr molecular markers and their application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Determination of corn SSR molecular markers and primers

[0032] 36 maize varieties were selected for SSR primer screening. Use denaturing polyacrylamide gel electrophoresis to select amplified sites with clear band patterns and high polymorphisms from 987 pairs of SSR primers (https: / / ftp.maizegdb.org / MaizeGDB / FTP / SSRs / ) Primers serve as optional primers. According to the molecular weight range and PIC value of the alleles of the optional primers in the 36 varieties, that is, each combination is composed of as many primers as possible, and the allele ranges of all primers in the group do not overlap, four primer sets are formed and combined. The primers with FAM, VIC, NED and PET fluorescent groups were synthesized respectively. See Table 1. Those skilled in the art can understand that other fluorophores including FAM, VIC, NED, PET can also be selected in the art to modify the four sets of primers in Table 1, as long as the primers in each group are label...

Embodiment 2

[0036] Example 2 Utilize the SSR molecular marker provided by the present invention to distinguish maize varieties

[0037] (1) Rapid extraction of DNA

[0038] DNA was extracted from maize seeds by alkaline boiling method. This method is quick and easy to operate, has no toxic and harmful reagents, is suitable for DNA preparation in the field of plant molecular biology, and is of great significance for greatly shortening the time of seed purity inspection and transgenic detection, improving detection efficiency, and reducing detection costs. The specific operation steps are as follows: take several corn seeds and place them in a 1.5mL centrifuge tube; add 400 μL NaOH (1M) to the centrifuge tube to ensure that the seeds are completely soaked, and bathe in boiling water for 5 min; add 200 μL Tris-HCl ( 1M, pH 8.0), in a boiling water bath for 1 min; add 200 μL TE buffer (pH 8.0), fully dissolve and set aside.

[0039] (2) Quality and quantity of DNA samples

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Abstract

The invention provides a corn SSR molecular marker group and application thereof. Seventeen markers are included in the corn SSR molecular marker group. The invention further provides a set of primercombination suitable for a capillary cataphoresis platform used for detecting the SSR molecular markers. By utilizing the seventeen SSR molecular markers, (1) an SSR marker fingerprint database of corn varieties can be constructed; (2), matrilineal traceability analysis of corn samples can be conducted; (3) authentication of the corn varieties and genetic diversity analysis can be conducted; (4) varieties of Demeiya No.1 and Denghai 18 of corns can be distinguished. By means of the application of the corn SSR molecular markers, the range of marker sites which can be used by corns on the levelof genetic group can be expanded; novel tools for conducting researches on the corn varieties, germplasm resource identification, genetic relationship evaluation, cytoplasmic genetic characteristics and the like are provided, and the application prospect is good.

Description

technical field [0001] The invention belongs to the technical field of crop molecular biology, and in particular relates to a group of corn SSR molecular markers used in capillary electrophoresis detection technology and applications thereof. Background technique [0002] Maize purity identification and variety differentiation are the key links in the maize seed quality control system. At present, the methods of maize purity identification and variety differentiation include morphological identification, isozyme or seed storage protein electrophoresis technology, etc. Among them, the seed storage protein electrophoresis technology has the advantages of simplicity, speed, accuracy, and low cost, and related technical standards formulated based on this technology are being widely used. However, due to the poor polymorphism of seed storage proteins among some maize varieties, or the bands appearing as non-complementary types, it is difficult to find characteristic bands by usi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 关海涛张瑞英张晓磊温洪涛黄盈莹
Owner 黑龙江省农业科学院农产品质量安全研究所