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Nucleic acid composition for detecting Datura ferox and application thereof

A nucleic acid composition and nucleic acid technology, applied in the fields of agriculture and plant inspection and quarantine

Active Publication Date: 2011-03-23
中华人民共和国上海海关
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no molecular level research results can be used to identify the spiny mandala (D.ferox)

Method used

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  • Nucleic acid composition for detecting Datura ferox and application thereof
  • Nucleic acid composition for detecting Datura ferox and application thereof
  • Nucleic acid composition for detecting Datura ferox and application thereof

Examples

Experimental program
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Effect test

Embodiment Construction

[0017] The technical solutions in the embodiments of the present invention will be clearly and completely described below.

[0018] 1. Sample DNA Extraction

[0019] The single seed is ground and pulverized, and extracted according to the following steps:

[0020] 1) Grind finely in liquid nitrogen and transfer to a centrifuge tube, add 500-800μL TES [100mM Tris (pH8.0), 10mM EDTA, 2% SDS]; add 50-100μg protein kinase K and mix well, place at 55℃~ Water bath at 60°C for 60-90 minutes, and mix gently several times during the period;

[0021] 2) To adjust the salt concentration, add 1 / 10 volume of 10% cetyltriethylammonium bromide (CTAB), and place in a water bath at 65°C for 10-20 minutes;

[0022] 3) Add an equal volume of SEVGA (chloroform: isoamyl alcohol = 24:1) to mix, and incubate at 0°C for 30 min;

[0023] 4) Centrifuge, take the supernatant, add 3-6 μL RNase A, and bathe in water at 37°C for 30 minutes to remove RNA;

[0024] 6) Add pre-cooled isopropanol, and plac...

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Abstract

The invention provides a nucleic acid composition for detecting Datura ferox, which comprises a nucleic acid primer F shown as SEQ ID No. 1, a nucleic acid primer R shown as SEQ ID No. 2 and a nucleic acid molecular probe P shown as SEQ ID No. 3 or a nucleic acid chain which is fully complementary with the nucleic acid primer F, the nucleic acid primer R and the nucleic acid molecular probe P. In the invention, by adopting a real-time fluorescent polymerase chain reaction (PCR) detection technology, the nucleic acid composition of the invention can complete detection within half of a working day, thus greatly shortening detection time and obviously improving detection sensitivity. The nucleic acid composition can be applied to the fields such as port inspection and quarantine, agricultural production, plant protection and the like, thus achieving the purpose of rapid and accurate detection and identification.

Description

technical field [0001] The present invention relates to the technical fields of agriculture and plant inspection and quarantine, in particular to a nucleic acid composition for quickly and accurately detecting D. ferox by using molecular biology detection technology, and adopting real-time fluorescent PCR detection technology, the composition It is applicable to fields such as port inspection and quarantine, agricultural production and plant protection. Background technique [0002] Datura L. belongs to Solanaceae. Datura poisoning incidents occur every year in the world. In addition, because many species of Datura are widely used in pharmacy and horticulture, many species of this genus are cultivated in large quantities all over the world and in my country. Widely distributed worldwide. In imported consignments such as soybeans, interception of seeds of the poisonous and noxious weed Datura spp. has been frequently reported, and seed identification is subject to confusion....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64C12N15/11
Inventor 印丽萍易建平魏莎莎傅怡宁薛华杰
Owner 中华人民共和国上海海关
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