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Erigeron breviscapus DNA polymorphism detection method and application thereof

A technology of DNA polymorphism and short scape flying canopy, which is applied in the measurement/testing of microorganisms, biochemical equipment and methods, etc., can solve the problem of less research work at the molecular level, and achieve the effect of promoting industrialization

Active Publication Date: 2010-12-15
YUNNAN SHIPURUI BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, the researches on Tcappa brevifolia have mainly focused on the extraction and identification of chemical components, physiological morphology, cultivation techniques and isoenzymes, while little research has been done on its molecular level.

Method used

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  • Erigeron breviscapus DNA polymorphism detection method and application thereof
  • Erigeron breviscapus DNA polymorphism detection method and application thereof
  • Erigeron breviscapus DNA polymorphism detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Total DNA Extraction

[0037] In this example, the total DNA of 40 experimental materials of Breviscapus breviscapus was extracted respectively, so as to be used for AFLP analysis of genetic diversity and molecular marker fingerprints of these materials.

[0038] Table 1 Name and source of experimental materials

[0039]

[0040]

[0041] The specific DNA extraction method is as follows: Take 0.2 g of the young leaves of the experimental material and add liquid nitrogen to grind to powder, add 1 ml of extraction buffer (containing 2% CTAB) and place it at 65 ° C for 1 h, during which it is constantly shaken. After taking out, centrifuge at 10000rpm for 10min (4°C). After the centrifugation, the supernatant was transferred to another clean centrifuge tube, and an equal volume of chloroform:isoamyl alcohol (24:1) was added and shaken constantly. After standing still for a while, centrifuge at 10,000 rpm for 10 min (4°C), take out the supernatant, add ...

Embodiment 2

[0043] Example 2 AFLP response

[0044] In this embodiment, the AFLP reaction is performed on the genomic DNA extracted in Example 1, which specifically includes the following steps:

[0045] 1) Digestion and ligation:

[0046] The genomic DNA extracted in Example 1 was subjected to EcoR1 / Mse1 double enzyme digestion, and the enzyme digestion system and conditions were as follows:

[0047] (1) EcoRI enzyme digestion system (20ul system / sample): EcoRI 1ul (10U / ul), 10× buffer 2ul, sample (total DNA) 5ul, ddH 2 O 12ul;

[0048] EcoR1 enzyme digestion conditions: 37°C for 2h, 65°C for 30min, stop the reaction;

[0049] (2) MseI enzyme digestion system (20ul system / sample): MseI 2ul (1U / ul), 10× buffer 2ul, sample (EcoRI digestion product) 15ul, ddH 2 O 1ul;

[0050] Mse1 enzyme digestion conditions: 65°C for 2 hours, 80°C for 30 minutes, stop the reaction;

[0051] Ligate the digested product with the synthetic linker fragment, the linker sequence is as follows:

[0052...

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Abstract

The invention provides an erigeron breviscapus DNA polymorphism detection method and application thereof. In the invention, an amplified fragment length polymorphism technology is used for erigeron breviscapus DNA polymorphism detection, an optimum primer combination is screened out, and reaction condition is optimized. The method can be effectively applied to the inheritance breeding, the resource protection, the introduction domestication, the breed optimization and the like of erigeron breviscapus.

Description

technical field [0001] The present invention relates to the DNA polymorphism detection method and application of the crocodile crocodile, specifically, the present invention adopts the technique of amplified fragment length polymorphism to detect the DNA polymorphism of the crocodile Background technique [0002] Amplified fragment length polymorphism (AFLP) is a DNA polymorphism detection technology developed on the basis of random amplified polymorphism (RAPD) and restriction fragment length polymorphism (RFLP). The high repeatability of RFLP technology and the simplicity and speed of RAPD technology do not require the preparation of probes like RFLP analysis, and the detection of genomic polymorphisms, like RAPD markers, does not need to know the sequence characteristics of the genome, while making up for the RAPD technology The defect of poor repeatability. Compared with other PCR-based marker technologies, AFLP technology can simultaneously detect a large number of sit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 鸭乔龙祥胡志祥谢德汝
Owner YUNNAN SHIPURUI BIOLOGICAL ENG
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