Method for fast identifying Qingxi black strain and Japanese strain in pelodiscus sinensis

A technology for Chinese soft-shelled turtles and strains, applied in the field of molecular biology, can solve problems such as molecular mechanism that needs to be further studied.

Active Publication Date: 2020-04-03
ZHEJIANG WANLI UNIV
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Problems solved by technology

At present, there is no report on the regulation of black turtle melanin synthesis, and the molecular mechanism of the black soft-shelled turtle’s traits needs to be further studied

Method used

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  • Method for fast identifying Qingxi black strain and Japanese strain in pelodiscus sinensis
  • Method for fast identifying Qingxi black strain and Japanese strain in pelodiscus sinensis

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Embodiment 1

[0014] The RNA samples from the abdominal skin tissue of the two strains of soft-shelled turtle extracted from cryopreservation were reverse-transcribed, and 2 μg of RNA was obtained according to TAKARA Primescript TM Ⅱ 1st strand cDNA synthesis Kit instructions for reverse transcription of cDNA. The reverse-transcribed cDNA was used as a template for ASP gene cloning, and PCR amplification was performed using Beijing Quanshijin HiFi Taq DNA Polymerase. Use 10-fold diluted cDNA as a template for gene cloning, prepare 50 μL reaction mixture, and add 5 μL 10×HiFi PCR Buffer II (containing Mg 2+ ), 1 μL cDNA template, 2 μL each primer, 4 μL 2.5 mM dNTP, 35.5 μL ddH 2 O and 0.5 μL HiFi Taq DNA Polymerase. After mixing evenly, amplify on a PCR instrument. The amplification conditions are 95°C for 2 minutes for pre-denaturation, 95°C for 30s for denaturation, 55°C for 30s (390bp) / 56.5°C for 30s (374bp) for annealing, and 72°C for 30s for 30 cycles. , 75°C for 5 minutes and finall...

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Abstract

The invention discloses a method for fast identifying a Qingxi black strain and a Japanese strain in pelodiscus sinensis. The method for fast identifying the Qingxi black strain and the Japanese strain in pelodiscus sinensis is characterized in that skin tissue RNA of to-be-identified pelodiscus sinensis is extracted, reverse transcription is conducted to obtain cDNA, with the cDNA as a template,PCR amplification is conducted, an amplification forward primer sequence is shown in SEQ ID No.1 in the description, a reverse primer sequence is shown in SEQ ID No.2 in the description, if a 374-bp amplification product can be obtained from amplification, the to-be-identified pelodiscus sinensis is the Japanese strain, and if no 374-bp amplification products can be obtained, the to-be-identifiedpelodiscus sinensis is the Qingxi black strain. The method for fast identifying the Qingxi black strain and the Japanese strain in pelodiscus sinensis can be used as a method for identifying the Qingxi black strain and the Japanese strain in pelodiscus sinensis for genetic breeding, especially breeding of a black character, of pelodiscus sinensis.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for quickly identifying the Qingxi soft-shelled turtle strain and the Japanese strain in the Chinese soft-shelled turtle. Background technique [0002] Chinese soft-shelled turtle (Pelodiscus sinensis) is an important famous and high-quality aquaculture species in my country. It has the nutritional characteristics of high protein and low fat, rich in various vitamins and trace elements, and has high nutritional value. The germplasm resources of Chinese soft-shelled turtle are the source of its industrial development. A new national aquatic variety recognized by the Fine Variety Approval Committee and announced by the Ministry of Agriculture. These two new varieties of soft-shelled turtle have their own unique external morphology and production performance characteristics. They are not only excellent varieties for breeding production, but also valuable germpl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/6888C12Q1/686C12Q2521/107Y02A40/81
Inventor 斯越秀尹尚军张丽丽钱国英
Owner ZHEJIANG WANLI UNIV
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