Molecular marker fingerprinting for identifying anoectochilus formosanus produced at Huoshan in Anhui and similar species thereof

A technology of fingerprints and molecular markers, which is applied in the field of fingerprints of Aurantia clematis, can solve problems such as time-consuming, high cost, and complicated procedures, and achieve the effect of simple steps, less sampling, and strong genotyping ability

Active Publication Date: 2013-04-24
安徽同济生生物科技有限公司
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  • Application Information

AI Technical Summary

Benefits of technology

This patented technique allows us to create genetic markers that help research scientists better study crops or other organisms' roots based on their natural characteristics such as size, shape, coloration, etc., without having them undergo physical damage during testing. It also helps predict how different types of medicine will work well together when mixed together due to similar properties from each type. Overall, this new discovery makes it possible to improve agricultural production efficiency while reducing costs associated therewith.

Problems solved by technology

The technical problem addressed in this patents is how to accurately distinguish various types of citrus crops like Xylonglavescenceceae (Coriidae). Existing methods require expensive equipment and complex processes, making them difficult to use outside China where they may affect healthy populations.

Method used

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  • Molecular marker fingerprinting for identifying anoectochilus formosanus produced at Huoshan in Anhui and similar species thereof
  • Molecular marker fingerprinting for identifying anoectochilus formosanus produced at Huoshan in Anhui and similar species thereof
  • Molecular marker fingerprinting for identifying anoectochilus formosanus produced at Huoshan in Anhui and similar species thereof

Examples

Experimental program
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Effect test

Embodiment

[0049] (1) Pretreatment of fresh clematis samples

[0050] Keep fresh clematis samples away from light for more than 6 hours to consume starch, take 10 grams of fresh clematis leaves, soak them in more than 50 grams of absolute ethanol twice, each time for 10 minutes, and wipe dry;

[0051] (2) Extraction of sample genomic DNA

[0052] The improved CTAB method was used to extract genomic DNA from leaves of A. clematis, and the operation was as follows:

[0053] 1) Put the pretreated leaves into a frozen ceramic mortar, add an appropriate amount of quartz sand, then add liquid nitrogen several times to fully grind to powder, and quickly transfer the leaf powder to a 10ml pre-cooled Eppendorf centrifuge tube with a medicine spoon When the height of the powder reaches 1 / 3 of the centrifuge tube, stop adding the powder;

[0054] 2) Add 4ml of 65°C preheated CTAB extraction buffer [2% (v / w) CTAB, 1.4M NaCl, 20mM EDTA, 100mM Tris-HCl (PH=8.0), 2% mercaptoethanol] into the centrifu...

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Abstract

The invention discloses a molecular marker fingerprinting for identifying the anoectochilus formosanus produced at Huoshan in Anhui and the similar species thereof. The molecular marker fingerprinting comprises the steps of preprocessing anoectochilus formosanus samples, extracting DNA of each sample genome, executing ISSR-PCR (inter-simple sequence repeat-polymerase chain reaction), executing agarose gel electrophoresis, building ISSR molecular marker fingerprint of each sample and using the ISSR molecular marker fingerprint for identifying the anoectochilus formosanus germplasm. The method provided by the invention saves time and economic cost, and can obtain the accurate and reliable identification fingerprint just by extracting the DNA of the anoectochilus formosanus plants, executing ISSR-PCR and executing agarose gel electrophoresis; the method identifies the species, which are easy to confuse in appearance, from the essence of inheritance, has the advantages of being simple, convenient and efficient, excellent in repeatability and high in resolution, and so on; and the method can identify the species in seeding stage of plants, and plays an important role in guaranteeing the accuracy and the stability of the Huoshan anoectochilus formosanus crude drug base source.

Description

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Claims

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Application Information

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Owner 安徽同济生生物科技有限公司
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