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Acyrthosiphon gossypii PCR (polymerase chain reaction) quick detection method based on specific SS-COI primers

A kind of cotton long tube aphid and detection method technology, which is applied in the field of molecular biology and achieves the effects of strong specificity, simple and fast operation and high accuracy

Active Publication Date: 2015-06-03
INST OF PLANT PROTECTION OF XINJIANG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, this method has not been applied in the detection of cotton aphid

Method used

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  • Acyrthosiphon gossypii PCR (polymerase chain reaction) quick detection method based on specific SS-COI primers
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  • Acyrthosiphon gossypii PCR (polymerase chain reaction) quick detection method based on specific SS-COI primers

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Embodiment Construction

[0027] The following specific examples are used to illustrate the present invention, but are not used to limit the scope of the present invention. Unless otherwise specified, the examples were operated according to the conventional molecular biology experimental conditions and the conditions suggested in the product instructions.

[0028] The first part, primers Ag22F / Ag22R for the amplification of Aphis gossypii

[0029] (1) Preparation of the cotton aphid genome

[0030] Aphids gossypii were collected in cotton fields, single-headed aphids were put into a 1.5mL centrifuge tube and ground, and the genome of single-headed aphids was extracted with the OMEGA Insect Genomic DNA Kit (OMEGA, USA). Store the DNA solution at -20°C for later use, and pipette 1 μL of the solution as a DNA template when performing PCR amplification.

[0031] (2) Synthesize specific SS-COI primers for detecting A. gossypii, the sequence is

[0032] Ag22F: 5'-TCTTAATTAATAATGGTACG-3' (SEQ ID No.1)

[...

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Abstract

The invention relates to an Acyrthosiphon gossypii PCR (polymerase chain reaction) quick detection method based on specific SS-COI primers. The primers are composed of a forward primer Ag22F and a reverse primer Ag22R. The primers only have specific amplification effect for Acyrthosiphon gossypii, the amplification product length is 260bp, and the DNA (deoxyribonucleic acid) minimum detection concentration is 0.170ng / mu L. When being used for Acyrthosiphon gossypii dominant natural enemy determination and food chain analysis in the field of plant protection, the method has the advantages of high specificity and high accuracy, and is convenient, quick and easy to operate.

Description

1. Technical field [0001] The invention belongs to the field of molecular biology, and relates to a PCR detection method, in particular to a PCR detection method based on the specific SS-COI primer of Aphid gossypii. 2. Background technology [0002] Cotton long tube aphid (Acyrthosiphon gossypii Mordvilko), also known as cotton netless long tube aphid, large cotton aphid, belongs to Homoptera, Aphididae. Harm cotton, camel thorn, bitter bean, broad bean, cowpea, mung bean, mallow mallow, licorice, etc. [0003] Cotton aphids are weakly clustered, scattered on the back of leaves, twigs and flower buds of cotton plants, and suck cotton juice with piercing-sucking mouthparts. The peak period of damage is in the bud stage, small chlorotic spots appear on the damaged parts, and the leaves curl slightly, which affects the development of cotton plants. When the insect population density is high, a large amount of nutrients in the cotton body are consumed, causing buds to fall off...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888
Inventor 王冬梅李海强刘建丁瑞丰李号宾徐遥戴爱梅阿克旦王剑
Owner INST OF PLANT PROTECTION OF XINJIANG ACADEMY OF AGRI SCI