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Ramie micro-satellite DNA Label

A microsatellite marker, ramie technology, applied in the field of ramie DNA molecular genetic markers, can solve the problems of limited developmental period, low accuracy, and inability to distinguish homozygous genotypes from heterozygous genotypes

Inactive Publication Date: 2006-08-09
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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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 ramie varieties and analysis of genetic relationships and the limitations of growing seasons and developmental periods is an urgent technology to be solved in the development of ramie research.

Method used

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  • Ramie micro-satellite DNA Label
  • Ramie micro-satellite DNA Label
  • Ramie micro-satellite DNA Label

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0021] 1: Ramie (CT / AG) n Construction of Microsatellite Enriched Library

[0022] (1) According to Guo Anping's method (see Guo Anping, 1997, China Mazuo, 19: 4-10) to extract ramie genomic DNA. Use restriction endonuclease Mse I digestion ramie genome DNA, digestion product is electrophoresis on 1.2% agarose gel, cuts out the DNA fragment of 300-1000bp size under ultraviolet light and uses gel recovery kit (purchased from Qiagen company) Purify the recovered digested product.

[0023] (2) base-complementary oligonucleotide PHC-1 (5'-AGATGGAATTCGTACACTCGT-3') and phosphorylated oligonucleotide PHC-2 (5'-phos-TAACGAGTGTACGAATTCCATCT-3') were mixed equimolarly, Denature at 95°C for 5 minutes and anneal at 60°C for 1 hour to form a linker with MseI sticky ends.

[0024] (3) Ligate the digested product and adapter with T4 DNA ligase. The ligated product was extracted once with an equal volume of phenol / chloroform / isoamyl alcohol (25:24:1), and the supernatant was collected by...

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Abstract

This invention relates to a DNA molecular genetic marker of ramee and provides 15 DNA sequences of the ramee micro-satellite sites and the primer sequences for expanding the sites and a method, in which, the sites are used in classifying its varieties, its genetic relation analysis and marker assistant breeding and is a reliable and effective molecular marker.

Description

technical field [0001] The invention relates to molecular marker technology, in particular to a DNA molecular genetic marker of ramie. Background technique [0002] Microsatellites (also known as simple repeats, SSRs) are short sequences consisting of 1 to 6 nucleotides that repeat multiple times in series and are widely distributed in eukaryotic genomes. There are conserved DNA sequences on both sides of the microsatellite sequence, and primers are designed according to the conserved sequences on both sides, and the sequence of the microsatellite site can be amplified by PCR. Since the number of repetitions of the repeating unit in the microsatellite is different, the length of the amplified microsatellite sequence is polymorphic (ie, SSRLP or SSR). Because of its rich polymorphism, high repeatability, co-dominant inheritance and other advantages, it is widely used in the construction of genetic maps, gene mapping, genetic relationship analysis, variety identification and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12P19/34C12N15/11
Inventor 揭雨成蒋彦波周建林张健
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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