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PCR (polymerase chain reaction) method and kit for simultaneously identifying specificities of Q-type bemisia tabaci and B-type bemisia tabaci

A technology of Bemisia tabaci and kit, applied in the field of specific PCR method and kit for simultaneous identification of Q-type and B-type Bemisia tabaci

Inactive Publication Date: 2013-05-08
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the primer pairs currently used in the molecular markers for B. tabaci and Q B. tabaci biotype identification can only identify a single biotype (Shatters et al., 2009)

Method used

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  • PCR (polymerase chain reaction) method and kit for simultaneously identifying specificities of Q-type bemisia tabaci and B-type bemisia tabaci
  • PCR (polymerase chain reaction) method and kit for simultaneously identifying specificities of Q-type bemisia tabaci and B-type bemisia tabaci
  • PCR (polymerase chain reaction) method and kit for simultaneously identifying specificities of Q-type bemisia tabaci and B-type bemisia tabaci

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Experimental program
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Embodiment 1

[0043] Embodiment 1 RAPD amplification screening specific random primer

[0044] 2.1 DNA extraction from single whitefly pest

[0045] Genomic DNA was extracted using a DNA extraction kit (Beijing Biotech Biotechnology Co., Ltd., solution type), and the method was carried out according to the kit instructions.

[0046] The specific operation is as follows: place a single adult Bemisia tabaci on the Parafilm membrane dripped with 10 μl of cell lysate (provided by the kit), use the bottom of a 0.2ml PCR tube as a homogenizer to grind fully, and use another 20 μl of lysate Wash the homogenizer twice, combine and mix; then transfer to a 1.5ml centrifuge tube with a micropipette; add proteinase K, 2 μl, mix well, 55 ° C, water bath for 4 hours or overnight; centrifuge for a few seconds, add RNase A2. Put 5 μl (10 μg / μl) in an oven at 37°C for 15-30 minutes; take out the above sample, let it cool to room temperature, add 25 μl of protein precipitation solution, and oscillate at hig...

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Abstract

The invention discloses a PCR (polymerase chain reaction) method and a kit for simultaneously identifying specificities of Q-type bemisia tabaci and B-type bemisia tabaci, and belongs to the technology of molecular biological detection. The PCR method comprises the steps of carrying out PCR amplification on DNA of the bemisia tabaci to be tested, and detecting PCR amplification product. The PCR method is characterized in that the primer pair adopted for PCR amplification is as follows: an upstream primer Bt-F:5'-TCT CAT CGG TGT CTTATT G-3', and a downstream primer Bt-R:5'-CTT CGG GTC GTT TCT CGT-3'; the detection of the PCR amplification product means that gel electrophoresis is carried out on the PCR amplification product to observe whether characteristic belt type appears; the characteristic belt type of the Q-type bemisia tabaci is a 507bp stripe; and the characteristic belt type of the B-type bemisia tabaci is 507bp and 660pb stripes. A kit is also provided on the basis of the method.

Description

technical field [0001] The invention belongs to biological detection technology, in particular to a specific PCR method and kit for simultaneously identifying Q-type and B-type Bemisia tabaci. Background technique [0002] Bemisia taabaci belongs to Hemiptera and belongs to the family Whitefly. It is widely distributed in all continents of the world. It causes serious harm to agricultural and horticultural production by directly feeding on plant juice, secreting honeydew to pollute plants, and spreading plant viruses. Bemisia tabaci is extremely invasive. Over the past two decades, whitefly has invaded many countries and regions around the world and replaced many local indigenous whiteflies, causing serious damage to many important crops around the world. Except In addition to the wilting of the leaves and even the plants caused by the sucking of the plant juice itself, the physiological disorder of the host plant, the most important thing is the large-scale outbreak of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王少丽王金娜张友军吴青君谢文徐宝云
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI