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Method for extracting total RNA (Ribonucleic Acid) of grapholita molesta adult antennae

A technology of pear borer and extraction method, which is applied in the field of extraction of total RNA from the antennae of small moth insects, can solve the problems of difficult extraction of high-quality antennae total RNA, and achieve the effect of solving degradation

Inactive Publication Date: 2012-08-15
NORTHWEST A & F UNIV
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Problems solved by technology

[0006] Aiming at the problem that it is difficult to extract high-quality total RNA from antennae of small moth insects such as adults of pear borer moth, the purpose of the present invention is to provide a simple and effective method for extracting total RNA from antennae of adults of pear borer

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  • Method for extracting total RNA (Ribonucleic Acid) of grapholita molesta adult antennae
  • Method for extracting total RNA (Ribonucleic Acid) of grapholita molesta adult antennae
  • Method for extracting total RNA (Ribonucleic Acid) of grapholita molesta adult antennae

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Embodiment Construction

[0014] The present embodiment provides a kind of method for extracting the total RNA of the tentacles of pear borer worm adults, which is implemented according to the following steps:

[0015] 1. First put 50 adult pear borer worms in a -20°C refrigerator and freeze them for 8 minutes, then place them on a piece of white paper, clamp the head of the insects with 12.5cm small medical tweezers, and gently lighten them with 12.5cm small medical scissors. Gently cut off the tentacles and immerse them in a capless 1.5ml centrifuge tube in liquid ammonia with a buoy with a diameter of 2cm and about 50ul of liquid nitrogen in the tube.

[0016] 2. After the tentacles of all adults are loaded into the centrifuge tube, take the centrifuge tube out from the liquid nitrogen and immediately add 125ul of total RNA extraction solution (Trizol, Shenzhen Jingmei Company), and use a micro electric tissue homogenizer (the homogenizer) Use a 1.5ml plastic grinding rod) to fully homogenize the ti...

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Abstract

The invention discloses a method for extracting total RNA (Ribonucleic Acid) of grapholita molesta adult antennae. The method comprises the following steps of: freezing grapholita molesta adults for coma under a low temperature condition, shearing a plurality of antennae by using small medical scissors and forceps, immediately soaking the antennae in a 1.5ml centrifugal tube in liquid nitrogen, taking the centrifugal tube out of liquid ammonia, immediately adding a small amount of RNA extraction liquid, fully homogenizing the extraction liquid by using a homogenizer, adding enough total RNA extraction liquid, blowing for a few times by use of a pipettor, transferring to the other new centrifugal tube, performing chloroform extraction according to a total RNA extraction method, and performing isopropanol and ethanol precipitation and separation, thus obtaining high-purity total RNA. The method is simple and convenient, and can be used for solving the problem about degradation of the RNA in the extraction process, so that the high-quality total RNA is obtained.

Description

technical field [0001] The invention relates to a method for extracting total RNA from antennae of small moth insects, in particular to a method for extracting total RNA from antennae of adults of the small moth moth. Background technique [0002] The small pear borer Grapholita molesta (Busck) is a worldwide important pest that seriously harms peaches, pears and other fruit trees. It is widely distributed in all provinces, cities and autonomous regions in my country except Tibet. In recent years, due to the adjustment of the industrial structure of agricultural production, the influence of factors such as changes in fruit cultivation models and management measures, the small pear borer has occurred in many places in my country year after year, which not only caused huge economic losses to the fruit industry, but also seriously Dampened the production enthusiasm of broad fruit growers. It has become one of the important factors restricting the further development of pears, p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 仵均祥张国辉许向利李怡萍
Owner NORTHWEST A & F UNIV
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