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Cleistogenes songorica LTR-RT molecular marker primer and application in cleistogenes songorica germplasm identification

A technology of molecular markers and cryptograms, applied in the biological field, achieves the effects of convenient operation, high sensitivity and accuracy, and low cost

Active Publication Date: 2019-10-25
LANZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the development of LTR-RT related markers and their application in variety identification have not been reported in the non-model plant Cryptography amansifolia

Method used

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  • Cleistogenes songorica LTR-RT molecular marker primer and application in cleistogenes songorica germplasm identification
  • Cleistogenes songorica LTR-RT molecular marker primer and application in cleistogenes songorica germplasm identification
  • Cleistogenes songorica LTR-RT molecular marker primer and application in cleistogenes songorica germplasm identification

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Experimental program
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Effect test

Embodiment

[0025] Concrete implementation steps flow chart of the present invention refers to figure 1 , including:

[0026] 1. Identification of material DNA extraction

[0027] Referring to the SDS method, the specific operation steps are as follows:

[0028] 1) Select 50 plump and healthy seeds and germinate them at 20°C for 72 hours by double-layer filter paper method;

[0029] 2) Mix the above seeds in a mortar and grind thoroughly, add 600 μL DNA buffer, and continue grinding and mixing. Transfer the grinding solution to a 2.0mL centrifuge tube;

[0030] 3) Transfer the centrifuge tube to a 65°C water bath for 10 minutes, and shake well 1-2 times during this process;

[0031] 4) Add 600 μL of a mixture of chloroform: isoamyl alcohol with a volume ratio of 24:1 in the fume hood, mix it upside down, centrifuge at 12,000 rpm for 10 min at room temperature, and transfer 360 μL of the supernatant into a 1.5 ml centrifuge tube;

[0032] 5) Add 36 μL of pre-cooled 3mol / L NaAc (pH=5.2...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a cleistogenes songorica LTR-RT (long-terminal repeat retrotransposons) molecular marker primer and application incleistogenes songorica germplasm identification. Specifically, 80 pairs of primer pairs and 23 kinds of cleistogenes songorica germplasm are used for distinguishing. According to the identification method, comparison and identification of the cleistogenes songorica germplasm can be completed within 4 hours by using few primer combinations, the advantages of being accurate, efficient, quick, low incost, convenient to operate and the like are achieved, and the sensitivity and accuracy are relatively high.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an LTR-RT molecular marker primer and its application in the germplasm identification of the genus Cryptospermum. Background technique [0002] Cleistogenes songorica (Cleistogenes songorica) is a perennial forage grass belonging to Gramineae. It mostly grows in arid grassland, desert or semi-desert sandy land, and is the construction and dominant species of desert grassland. It has strong drought tolerance and can grow in areas with an annual rainfall of 120 mm. It has strong cold resistance and heat resistance, is more resistant to trampling, and has a long green period. It can be used not only for animal husbandry production, but also for general lawn plants. It plays an important role in the stability of the ecosystem in the arid, semi-arid and arid desert regions of Northwest my country, and has important ecological and genetic values. There are more than 20 species of Cryptosper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 张吉宇马甜甜李洁吉赛尔吴凡闫启张正社
Owner LANZHOU UNIVERSITY
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