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Method for direct polymerase chain reaction (PCR) adopting litopenaeus vannamei hemolymph

A technology of hemolymph and white shrimp, applied in the field of molecular biology, can solve the problems of high cost of PCR technology, time-consuming and labor-intensive parental injury, etc., and achieve the effect of preventing hemolymph coagulation, low cost and less extraction amount

Pending Publication Date: 2019-04-19
南通龙洋生物科技有限公司
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Problems solved by technology

[0004] The purpose of the present invention is to solve the problems of the existing Penaeus vannamei PCR technology, which are costly, time-consuming and labor-intensive, and cause great damage to the parents, and provide a method for direct PCR using the hemolymph of Penaeus vannamei

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  • Method for direct polymerase chain reaction (PCR) adopting litopenaeus vannamei hemolymph
  • Method for direct polymerase chain reaction (PCR) adopting litopenaeus vannamei hemolymph
  • Method for direct polymerase chain reaction (PCR) adopting litopenaeus vannamei hemolymph

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Embodiment Construction

[0021] In order to describe in more detail, the present invention will be described below through a specific example and in conjunction with the accompanying drawings.

[0022] Get a fresh and alive Penaeus vannamei, carry out vannamei by the method said in the above-mentioned technical scheme MSTN The PCR of gene, concrete implementation steps are as follows:

[0023] 1. Anticoagulant preparation

[0024] Weigh 2.92g of EDTA, 20.67g of glucose, 19.25g of sodium chloride, and 8.22g of sodium citrate, add them into 1L of ultrapure water, mix and dissolve them thoroughly, and filter them with a 0.22um bacterial filter for later use.

[0025] 2. Hemolymph collection and pretreatment

[0026] a. Use a 1ml syringe to aim at the basement membrane at the junction of the sixth somite and telson of the prawns at an angle of 45°, insert 4 mm into the basement membrane, and extract 20ul of hemolymph.

[0027] b. Inject 20ul of the extracted hemolymph into a 200ul PCR tube, add 30ul o...

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Abstract

The invention discloses a method for a direct polymerase chain reaction (PCR) adopting litopenaeus vannamei hemolymph. The method comprises the steps of 1, anticoagulant preparation; 2, hemolymph collection and pretreatment; 3, PCR treatment. The method has the advantages of being easy to operate, low in cost, safe and free of toxins, the complex step of extracting DNA is omitted, harm to parent prawns is avoided, and the modern biological requirements are met.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a PCR method for prawns. Background technique [0002] Penaeus vannamei, because of its high yield, short breeding cycle, high meat yield, and delicious taste, has developed into a worldwide economical crustacean farming species. Since the introduction of Penaeus vannamei, it has actively cultivated excellent new species with independent intellectual property rights. However, the cultivation of domestic prawn varieties relies on traditional breeding and hybridization methods, and the cultivated varieties cannot be compared with foreign imported varieties. Therefore, it is the only way for the long-term development of Penaeus vannamei in China to use the means of molecular assisted marker breeding to breed safe, high-efficiency and controllable traits of hybrid offspring. In this process, it is very important to quickly obtain the genetic data of large samples, and the traditiona...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12Q1/686C12Q2527/125
Inventor 陈义培钱晓明赵岩岩闫兵兵张琳琳
Owner 南通龙洋生物科技有限公司
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