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Set of paddy rice SSR molecular markers suitable for capillary electrophoresis detection technology, and applications thereof

A capillary electrophoresis and molecular labeling technology, applied in the field of molecular biology of crops, can solve the problems of difficult unified processing of different batches of reaction data, difficult to accurately identify different allelic variations, non-automation, etc., and achieve accurate fragment size and calculated fragment size. , the effect of improving efficiency

Active Publication Date: 2018-08-07
黑龙江省农业科学院农产品质量安全研究所
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AI Technical Summary

Problems solved by technology

The traditional SSR operation uses polyacrylamide gel electrophoresis combined with some other biotechnologies to analyze gene polymorphisms. These methods are non-automated and time-consuming, and it is difficult to accurately identify different allelic variants, and it is difficult to uniformly process different batches of reaction data.

Method used

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  • Set of paddy rice SSR molecular markers suitable for capillary electrophoresis detection technology, and applications thereof
  • Set of paddy rice SSR molecular markers suitable for capillary electrophoresis detection technology, and applications thereof
  • Set of paddy rice SSR molecular markers suitable for capillary electrophoresis detection technology, and applications thereof

Examples

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Effect test

Embodiment 1

[0032] Example 1 Determination of rice SSR molecular markers and primers

[0033] 36 rice varieties were selected for SSR primer screening. Using denaturing polyacrylamide gel electrophoresis, 800 pairs of SSR primers (http: / / www.gramene.org / ) were used to select primers with amplified sites, clear band patterns, and high polymorphism 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 labeled with the same fl...

Embodiment 2

[0037] Embodiment 2 uses the SSR molecular marker provided by the present invention to distinguish rice varieties

[0038] (1) Rapid extraction of DNA

[0039] DNA was extracted from rice 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 a few rice 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.

[0040] (2) Quality and quantity of DNA samples

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Abstract

The invention provides a set of paddy rice SSR molecular markers suitable for capillary electrophoresis detection technology, and applications thereof. The set of paddy rice SSR molecular markers suitable for capillary electrophoresis detection technology comprises 13 SSR molecular markers. The invention also provides a primer combination which is used for detecting the SSR molecular markers and is suitable for capillary electrophoresis platforms. The 13 SSR molecular markers can be used for 1, construction of a paddy rice variety SSR labeled fingerprint database; 2, paddy rice maternal line source tracing analysis; 3, paddy rice variety identification and genetic diversity analysis, and 4, distinguishing of Beidao 7 and Longjing 29. Applications of the paddy rice SSR molecular markers arecapable of widening paddy rice available marker-site range in genome level, and providing novel tools in paddy rice variety and germplasm resources identification, genetic relationship evaluation, and cytoplasm hereditary character study, and the application prospect is promising.

Description

technical field [0001] The invention belongs to the technical field of crop molecular biology, and in particular relates to a group of rice SSR molecular markers suitable for capillary electrophoresis detection technology and applications thereof. Background technique [0002] Rice is one of the most important food crops in my country, and its sown area and total output both rank first in food crops. About 60% of the country's population feeds on rice. my country's rich rice seed resources are a major advantage in rice research and production. After more than 80 years of breeding practice, more than 5,000 rice varieties have been bred and applied in production. The identification of rice varieties, genetic diversity analysis and the construction of DNA fingerprint database can objectively and comprehensively understand the current status of rice varieties, which is of great significance for variety management, variety selection and germplasm resource collection and protecti...

Claims

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Application Information

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