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Core primer composition for Brassica SSR (simple sequence repeats)

A combination of primers and Brassica technology, applied in the field of molecular genetics, can solve problems such as waste, inaccurately grasping the overall effect of markers, time-consuming and labor-intensive problems, and achieve the effects of improving utilization, speeding up molecular marker work, and reducing costs

Inactive Publication Date: 2011-08-17
SOUTHWEST UNIVERSITY
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  • Abstract
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AI Technical Summary

Problems solved by technology

When we use SSR primers as markers, the usual practice is to screen all known SSRs and then select the primers we need. As a result, we will not be able to grasp the overall effect of the markers, resulting in a large waste of time and effort.

Method used

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  • Core primer composition for Brassica SSR (simple sequence repeats)
  • Core primer composition for Brassica SSR (simple sequence repeats)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0053] Selection of core primers for each chromosome of Brassica and establishment of multiplex PCR system

[0054] 1. Download 36 linkage maps of Brassica crops from the NCBI website, organize the SSR primers and marker information on each linkage group of each map, including the sequence of the primers, the position on the chromosome and the name of the linkage map;

[0055] 2. Selection of core SSR primers: Select primers that exist on more than 3 genetic maps, the annealing temperature range between primers is within 5°C, and are equidistantly distributed on the linkage group for combination. Select 10 primers for each linkage group. Primers for the core SSR;

[0056] 3. Multiplex PCR system and program optimization: multiplex PCR system: Buffer (10×) 2.5ul, primer (10umol / L) 2ul, dNTPs (10mM) 0.5uL, Taq DNA polymerase (5U / ul) 0.25ul, DNA template (50-100ng / ul) 2.0 uL, total 25ul reaction volume. The multiplex PCR program was: pre-denaturation at 94°C for 5 min; denatura...

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Abstract

The invention discloses a core primer composition for Brassica SSR (simple sequence repeats) and belongs to the technical field of molecular inheritance. The core primer composition comprises 190 core SSR primer pairs for 19 chromosomes in Brassica A and C genomes; and 10 SSR primer pairs are uniformly distributed on each chromosome, which have high polymorphism levels, substantially consistent annealing temperature and clear amplification band patterns. The core SSR primer composition is applied to increasing the molecular marking efficiency by 3 to 5 times without increasing the working costs, and is particularly suitable for the identification, the genetic diversity analysis, the nullisome and abnormal chromosome individual identification and the seed purity determination of the Brassica germplasm resources. The core SSR primer composition has the advantages of simpleness in operation, high stability and good reproducibility.

Description

technical field [0001] The invention relates to the technical field of molecular genetics, in particular to a combination of Brassica core SSR primers. Background technique [0002] Molecular markers are genetic markers based on nucleotide sequence variation in genetic material between individuals, and are a direct reflection of genetic polymorphism at the DNA level. Due to the large information content, DNA has a high degree of genetic stability within the same species, and is not affected by external environmental factors, individual developmental stages, and differences in organs and tissues. Molecular marker technology is a new marker technology following morphological markers, biochemical immunogenetic markers and cytogenetic markers, and has achieved a leap from phenotype selection to genotype selection. With the development of molecular biology technology, DNA molecular marker technology has been widely used in the breeding and production of corn, soybean, chicken, p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11
Inventor 付东辉魏丽娟李加纳高才华
Owner SOUTHWEST UNIVERSITY
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