Bluish dogbean micro satellite DNA label

A technology of DNA markers and microsatellite markers, which is applied in the field of DNA molecular genetic markers of kenaf, can solve the problems of inability to distinguish homozygous genotypes from heterozygous genotypes, growth seasons, developmental period restrictions, and low accuracy.

Inactive Publication Date: 2007-02-28
HUNAN NORMAL UNIVERSITY
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Problems solved by technology

However, RAPD markers are dominant in most cases and cannot distinguish homozygous genotypes from heterozygous genotypes
How to solve the problems of low accuracy in the classification of kenaf varieties and genetic relationship analysis, and the problems of being limited by the growing season and development period are the technologies that need to be solved urgently in the development of kenaf research.

Method used

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  • Bluish dogbean micro satellite DNA label
  • Bluish dogbean micro satellite DNA label
  • Bluish dogbean micro satellite DNA label

Examples

Experimental program
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Effect test

Embodiment Construction

[0012] 1: Kenaf (CT / AG) n Construction of microsatellite enrichment library

[0013] (1) Extract kenaf genomic DNA according to the method of Guo Anping (see Guo Anping, 1997, Chinese Ma Zuo, 19: 4-10). The kenaf genomic DNA was digested with restriction enzyme Mse I, and the digested product was electrophoresed on a 1.2% agarose gel. DNA fragments with a size of 300-1000 bp were cut under ultraviolet light and a gel recovery kit (purchased from Qiagen) ) Purify the recovered digested product.

[0014] (2) Mix the base complementary oligonucleotide PHC-1 (5'-AGATGGAATTCGTACACTCGT-3') and phosphorylated oligonucleotide PHC-2 (5'-phos-TAACGAGTGTACGAATTCCATCT-3') equimolarly, Denaturation at 95°C for 5 minutes and annealing at 60°C for 1 hour to form a linker with MseI sticky ends.

[0015] (3) Connect the digested product and the linker with T4 DNA ligase. The ligation product was extracted once with an equal volume of phenol / chloroform / isoamyl alcohol (25:24:1), and the supernatant...

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Abstract

A DNA molecular genetic marker of kenaf is disclosed. The DNA sequences of 5 polymorphic micro-satellite DNA markers are separated and cloned. Their primer sequences are amplified. Said micro-satellite DNA markers can be used for classifying the varieties of kenaf, discriminating the genetic relation and culturing new variety.

Description

Technical field [0001] The invention relates to molecular marker technology, in particular to a DNA molecular genetic marker of kenaf. Background technique [0002] Microsatellites (also known as simple repetitive sequences, SSR) are a piece of DNA composed of a short sequence of 1 to 6 nucleotides repeated multiple times in tandem, and are widely distributed in the eukaryotic genome. There are conservative DNA sequences on both sides of the microsatellite sequence, and primers are designed according to the conservative sequences on both sides of the microsatellite sequence, and the sequence of the microsatellite site can be amplified by PCR. Due to the different number of repetitions of the repeating unit in the microsatellite, the length of the amplified microsatellite sequence shows polymorphism (ie SSRLP or SSR). Because of its rich polymorphism, high reproducibility, codominant inheritance, etc., it is widely used in genetic map construction, gene mapping, genetic relationsh...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12Q1/68C07H21/04
Inventor 周建林揭雨成蒋彦波张健
Owner HUNAN NORMAL UNIVERSITY
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