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Method of identifying varieties of robinia pseudoacacia based on fluorescent SSR finger-prints of capillary electrophoresis

A capillary electrophoresis and fingerprint technology is applied in the field of identifying Robinia pseudoacacia species based on capillary electrophoresis fluorescence SSR fingerprints, and achieves the effects of convenient operation, less environmental impact, and rapid identification.

Active Publication Date: 2017-12-22
SHANDONG FOREST SCI RES INST +2
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Problems solved by technology

These SSR markers have not been used in the construction of Robinia pseudoacacia fingerprints

Method used

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  • Method of identifying varieties of robinia pseudoacacia based on fluorescent SSR finger-prints of capillary electrophoresis
  • Method of identifying varieties of robinia pseudoacacia based on fluorescent SSR finger-prints of capillary electrophoresis
  • Method of identifying varieties of robinia pseudoacacia based on fluorescent SSR finger-prints of capillary electrophoresis

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Embodiment 1

[0030] 1. Experimental materials:

[0031] In this experiment, 12 clones of Robinia pseudoacacia were used as test materials. R.pseudoacacia LuCi 1, R.pseudoacacia LuCi 2, R.pseudoacacia LuCi 7, R.pseudoacacia LuCi 8, R.pseudoacacia LuCi 9 ( R.pseudoacacia LuCi 9), Luci 10(R.pseudoacaciaLuCi 10), Luci 11(R.pseudoacacia LuCi 11), Luci 13(R.pseudoacacia LuCi 13), Luci 14(R.pseudoacacia LuCi 14), Luci 28 (R. pseudoacacia LuCi 28), Luci 32 (R. pseudoacacia LuCi 32), Luci 36 (R. pseudoacacia LuCi 36).

[0032] The materials were collected from Daqingshan Forest Farm, Fei County, Shandong Province.

[0033] In May 2016, the young leaves were collected, dried on silica gel, and set aside.

[0034] 2. Genomic DNA extraction:

[0035] Take the young leaves of Robinia pseudoacacia clones, and use the plant tissue genomic DNA extraction kit (magnetic bead method) (Cat. No. PTED-6030) of Inrich Biochemical Technology (Shanghai) Co., Ltd. to extract according to the instructions.

[0...

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Abstract

The invention discloses a method of identifying varieties of robinia pseudoacacia based on fluorescent SSR finger-prints of capillary electrophoresis. The construction method comprises the following steps: (1) extracting total DNA of robinia pseudoacacia; (2) performing PCR amplification by means of SSR primers; (3) performing capillary electrophoresis on a PCR amplification product; and (4) constructing finger-print, wherein the four pairs of disclosed SSR primers in the step (2) are used to perform PCR amplification of DNA of robinia pseudoacacia, the banding pattern results detected by capillary electrophoresis on the obtained product are combined to form the SSR finger-print of each variety or a clone, and the clone is then differentiated according to difference of the finger-prints. The method disclosed by the invention can provide a reliable ground for variety identification, new variety breeding and genetic improvement of robinia pseudoacacia from the molecular level quickly, accurately and efficiently, thereby providing reference for conservation, evaluation and utilization of genetic resources of Chinese robinia pseudoacacia.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying black locust varieties based on capillary electrophoresis fluorescence SSR fingerprints. Background technique [0002] Robinia pseudoacacia L. is a broad-leaved tree of the genus Robinia in the leguminous family (Leguminosae). It is resistant to drought, barrenness, mild salinity, pollution, and natural renewal. Pioneer tree species, and has ornamental, timber, honey source and feeding value. Robinia pseudoacacia, as an alien species, has been cultivated in my country for nearly 120 years. The source of Robinia pseudoacacia clones cultivated in various regions of China is unclear, the genetic relationship is unknown, and there is little information on genetic diversity. The phenomenon of homonyms and synonyms occurs from time to time, and a system is urgently needed. Carrying out genetic diversity analysis and fingerprint construction of Robinia pseudoacacia ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125C12Q2563/107
Inventor 毛秀红荀守华郑勇奇孙百友张元帅韩丛聪董玉峰
Owner SHANDONG FOREST SCI RES INST
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