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Physalis pubescens molecule-specific marker primers and identification method using primers

A technology for labeling primers and specificity, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc. It can solve the problems that there are no molecular specific labels, etc., and achieve simple methods, high sensitivity, and use less effect

Active Publication Date: 2017-02-15
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there is no research report on the application of molecular specific markers in the identification of Physalis pilosula

Method used

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  • Physalis pubescens molecule-specific marker primers and identification method using primers

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Effect test

Embodiment 1

[0018] Example 1: Development and Design of Molecular-Specific Labeling Primers for Physalis Physalis

[0019] 1. Genomic DNA Extraction

[0020] Cut 100 mg of fresh leaves of Physalis plant samples and put them into a mortar. The samples include Physalis (collected from 1: Tangshan, Hebei; -6: Hangzhou, Zhejiang, 7: Huanggang, Hubei, 8: Honghe, Yunnan, 9: Nanjing, Jiangsu, 10: Taizhou, Zhejiang, 11: Wenzhou, Zhejiang, 12: Jinhua, Zhejiang, 13: Dezhou, Shandong), Physalis (14: Shenyang, Liaoning, 15-16: Dandong, Liaoning, 17: Jinan, Shandong) and Maophysalis (18: Shenyang, Liaoning, 19: Zhaodong, Heilongjiang, 20: Changchun, Jilin). Immediately add liquid nitrogen and grind to powder, then use UNIQ-10 column type plant genomic DNA extraction kit (purchased from Shanghai Sangong Bioengineering Co., Ltd.) to extract genomic DNA, and the resulting DNA is detected by electrophoresis with 0.8% agarose gel, and used The concentration was detected by ultraviolet spectrophotometer a...

Embodiment 2

[0023] Example 2: PCR Amplification and Electrophoretic Detection of Molecular-Specific Marker Primers

[0024] The designed and synthesized primers ST3MSJF / ST3MSJR were used for PCR detection of different Physalis plant samples (see the description for details).

[0025] PCR reaction system (total volume 20μl): 2μl 10×Buffer, 2μl MgCl 2 (25mM), 0.8μl dNTPs (10mM), 1μl primer ST3MSJF (10μM), 1μl primer ST3MSJR (10μM), 1μl template DNA (50ng / μl), 0.5μl Taq enzyme (2U / μl), 11.7μl ddH 2 O.

[0026] PCR reaction program: pre-denaturation at 94°C for 5 min; 35 cycles (denaturation at 94°C for 50 s, annealing at 56°C for 50 s, extension at 72°C for 1.5 min); extension at 72°C for 10 min.

[0027] Use 1.5% agarose gel to detect the PCR product by electrophoresis, the electrophoresis picture is as follows figure 1 shown, from figure 1 (In the figure, channel M is the DNA molecular weight standard marker DL2000; channels 1-4: P. minima; 5-13: P. angulata; 14-17: P. alkekengi var. f...

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Abstract

The invention relates to molecule-specific marker primers for identifying Physalis pubescens and a method for quickly identifying Physalis pubescens by using the specific primers. The sequences of the primers are as follows: forward primer ST3MSJF: 5'-ATTGATAGGGTAAACCATG-3; and reverse primer ST3MSJR: 5'-CTGTAATAAAACAAAAGCG-3'. The specific primers ST3MSJF / ST3MSJR can be utilized to quickly identify the Physalis pubescens by a conventional PCR (polymerase chain reaction) process. The primers have the advantages of low consumption of laboratory samples, accurate results and high sensitivity; and the ST3MSJF / ST3MSJR are the specific amplification primers for Physalis pubescens, and if the sample is any other Physalis plant sample, the result is negative reaction. The method is simple and convenient; and by adopting the conventional PCR technique to perform detection, the consumed time is short, and the method can be completed within half a day.

Description

technical field [0001] The invention belongs to the technical field of identifying Physalis pubescens L. by means of molecular biology methods, relates to a specific marker primer for identifying Physalis pubescens L., and a method for quickly identifying Physalis pubescens by using the specific primer . Background technique [0002] Physalis pubescens L., aliased as yellow mushrooms and mushrooms, is an annual herbaceous plant of the genus Physalis in the family Solanaceae. It is native to the Americas. It is cultivated or wild in Jilin, Heilongjiang, Liaoning and the southeastern edge of eastern Inner Mongolia. It mostly grows in grasslands or beside roadsides. Physalis edulis is a very important medicinal and edible plant. As a folk traditional Chinese medicine, Physalis edulis was first included in "Shen Nong's Materia Medica", and it was listed as a middle grade. Its calyx-bearing fruit or whole plant is called lantern grass. It has a sour and flat taste and enters th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 冯尚国焦凯丽姜梦莹史钰军王慧中
Owner HANGZHOU NORMAL UNIVERSITY
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