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A kind of nucleotide sequence, specific primer and method for identifying bitter

A nucleotide sequence and specific technology, which can be used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. , the effect of short time

Active Publication Date: 2018-07-31
浙江美新控股有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The morphological characteristics of Physalis and other Physalis plants such as P. minima, P. alkekengi and P. pubescens are very similar (including stems, leaves, flowers and fruits). Therefore, using The traditional morphological classification method is difficult to quickly and accurately identify Sophora and other plants similar to Physalis
Therefore, in the past, it often happened that other Physalis plants were mistaken for Alopecia oleifera and used, which brought difficulties to the safe use and protection of Alopecia oleracea medicinal resources

Method used

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  • A kind of nucleotide sequence, specific primer and method for identifying bitter
  • A kind of nucleotide sequence, specific primer and method for identifying bitter
  • A kind of nucleotide sequence, specific primer and method for identifying bitter

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1: Preparation of the characteristic nucleotide sequence of Solanum oleifera

[0023] 1. Extraction of Genomic DNA

[0024] Cut Physalis plant samples (see Table 1) and put 100mg of fresh leaves into a mortar, immediately add liquid nitrogen to grind to powder, and then use UNIQ-10 column plant genomic DNA extraction reagent from Shanghai Sangon Bioengineering Co., Ltd. Genomic DNA was extracted from the box, and the resulting DNA was detected by electrophoresis with 0.8% agarose gel, and the DNA concentration was detected with an ultraviolet spectrophotometer, and diluted to 50 ng / μl.

[0025] Table 1. Physalis samples used in invention experiments

[0026]

[0027]

[0028] 2. SCoT–PCR reaction, electrophoresis detection

[0029]SCoT general primer 3 (5'-CAACAATGGCTACCACCG-3') was used for PCR amplification. The primer was synthesized by Shanghai Sangon Bioengineering Co., Ltd. The reaction system (total volume 20 μl) was: 10×Buffer 2 μl, MgCl 2 (25mM...

Embodiment 2

[0034] Example 2: Preparation of the specific primer ST3KZF / ST3KZR, PCR amplification, electrophoresis detection

[0035] On the basis of obtaining the specific nucleotide sequence of Sophora japonica, use Primer Primer 5.0 software to design the nucleotide sequence of ST3KZF / ST3KZR (respectively shown in SEQ ID NO.2, SEQ ID NO.3), primer Synthesized by Shanghai Sangon Bioengineering Co., Ltd. Then, the designed and synthesized primers ST3KZF / ST3KZR are used to amplify and detect different Physalis plant samples (see the accompanying drawings for details).

[0036] The PCR reaction system (total volume 20μl) is: 10×Buffer 2μl, MgCl 2 (25mM) 2μl, dNTPs (10mM) 0.8μl, primer ST3KZF (10μM) 1μl, primer ST3KZR (10μM) 1μl, template DNA (50ng / μl) 1μl, Taq enzyme (2U / μl) 0.5μl, ddH 2 O 11.7 μl.

[0037] The PCR reaction program was pre-denaturation at 94°C for 5 min; 35 cycles (denaturation at 94°C for 50 s, annealing at 65°C for 50 s, extension at 72°C for 1.5 min); and finally ext...

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Abstract

The invention relates to a nucleotide sequence, a specific primer and a method for identifying physalis angulata. The nucleotide sequence for identifying physalis angulata disclosed by the invention is shown as SEQ ID NO.1. The specific primer for identifying physalis angulata has an upstream sequence ST3KZF shown as SEQ ID NO.2 and a downstream sequence ST3KZR shown as SEQ ID NO.3. The physalis angulata is identified by a conventional PCR method by utilizing the specific primer ST3KZF / ST3KZR. The sample amount is small, and the whole operation can be finished by virtue of a small amount of samples; the accuracy and sensitivity are high, the ST3KZF / ST3KZR refers to a specific amplification primer of the physalis angulata, and if the samples refer to other physalis plants, a negative reaction presents. The method is simple and convenient, the PCR technology is adopted for detecting, the time consumption is low, and the operation can be finished within 3-5 hours.

Description

technical field [0001] The invention belongs to the technical field of using molecular biological methods to identify bitter beetroot, and relates to a nucleotide sequence for identifying bitter beetroot, specific primers and a method thereof. Background technique [0002] Physalis angulata L. is an annual herb of the genus Physalis of the family Solanaceae, distributed in tropical and subtropical regions of the world, often growing at an altitude of 500-1500 meters. In my country, it is mainly distributed in East China, Central China, South China and Southwest China. As a traditional medicinal material, for centuries, bitter fennel has been used as medicine with its fruit, root or whole herb, which can be used for the treatment of many diseases. its description. Its nature and flavor are bitter and cold, and it is mainly used to treat sore throat, mumps, acute and chronic bronchitis, lung abscess, dysentery, dysuria, and external application to treat impetigo. Modern med...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/156
Inventor 王慧中冯尚国姜梦莹焦凯丽史钰军
Owner 浙江美新控股有限公司
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