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Flax micro-satellite DNA mark

A technology of DNA markers and microsatellite markers, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., and can solve the problems that restrict the research and application of microsatellite markers

Inactive Publication Date: 2010-03-10
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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AI Technical Summary

Problems solved by technology

This seriously restricts the research and application of microsatellite markers in flax genetics and molecular biology

Method used

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  • Flax micro-satellite DNA mark
  • Flax micro-satellite DNA mark
  • Flax micro-satellite DNA mark

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0031] 1. Construction of flax microsatellite enrichment library.

[0032] (1) Genomic DNA of flax was extracted using improved CTAB method (see Deng Xin et al., 2007, China Hemp Industry Science, 29(4): 184-188) using the flax variety Baihua seedlings as materials. 10 μg of flax genomic DNA was digested with restriction endonuclease Mse I, and the digested product was detected by 1.5% agarose gel electrophoresis, and a DNA fragment of 300-1 500 bp was recovered with an agarose gel recovery kit (TIANGEN).

[0033] (2) The base-complementary oligonucleotide primer A (5'-AGATGGAA TTCGTACACTCGT-3') and primer B (5'-TAACGAGTGTACGAATTCCATCT-3') were mixed equimolarly, denatured at 95°C for 5min, annealed at 60°C for 1h, Forms adapters with sticky ends.

[0034] (3) Digest the digested product and adapter with T 4 DNA ligase ligation. The ligated product was extracted once with phenol / chloroform / isoamyl alcohol (25:24:1), and the supernatant was collected by centrifugation. Add ...

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Abstract

The invention discloses a flax micro-satellite DNA mark and provides a DNA sequence of 25 flax micro-satellite loci and an enlarging method for enlarging the primer sequence of the 25 flax micro-satellite loci. The 25 flax micro-satellite loci are used for classifying flax species, analyzing the genetic affinity and marking auxiliary culturing. With favorable repetitiveness, the flax micro-satellite DNA mark is a reliable and effective molecular mark.

Description

technical field [0001] The invention relates to a flax microsatellite DNA marker, which is suitable for flax variety classification, genetic relationship identification and auxiliary breeding. Background technique [0002] Microsatellite (Microsatel lite), also known as simple sequence repeat (SSR) or short tandem repeat (short tandem repeat, STR), is a repeat unit with a few nucleotides (1-6) Repeat DNA sequence in tandem. Microsatellites are widely used in genetic map construction, gene mapping, marker-assisted breeding, family analysis, variety identification, and evolutionary analysis due to their high level of heterozygosity and mutation rate, parental inheritance patterns, etc. And so on. Such as using microsatellite markers to study the genetic relationship between cherry varieties (Ai Chengxiang et al., 2007, Acta Fruit Tree, 24 (4): 460465), the patent application for pig breed classification with microsatellite DNA markers "suitable for pig breed classification ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 陈信波邓欣龙松华王玉富
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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