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Primers and method for distinguishing Thai marble goby fries from local marble goby fries

A technology of fish fry and primer sequence, applied in the field of aquaculture, can solve problems such as improper post-processing, and achieve the effects of low cost, simple and convenient operation, and assistance in aquaculture production

Inactive Publication Date: 2013-10-09
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the breeder accidentally mixes up the species, it will inevitably lead to improper handling in the later stage. As a result, the economic loss to the breeder will be immeasurable

Method used

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  • Primers and method for distinguishing Thai marble goby fries from local marble goby fries
  • Primers and method for distinguishing Thai marble goby fries from local marble goby fries

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Embodiment Construction

[0023] 1. Extraction of genomic DNA (cloud-spotted pond snakehead population, river sand pond snakehead population):

[0024] 1) Take 0.1~0.3g of the caudal fin of the experimental fish and add 450μL DNA extraction buffer (10 mmol / L Tris-HCl, 5 mmol / L EDTA, 75 mmol / L NaCl), mix well, and add SDS and proteinase K to The final concentrations were 0.5% and 200 μg / mL, respectively, and then digested overnight in a 55℃ water bath.

[0025] 2) Add an equal volume of saturated phenol, phenol / chloroform / isoamyl alcohol (25:24:1), chloroform / isoamyl alcohol (24:1) and extract once each. After each extraction was gently mixed for 10 minutes, centrifuged at 4°C and 12000 rpm for 20 minutes, and the supernatant was taken.

[0026] 3) Add 2.5 mol / L NaCl (final concentration of 0.2 mol / L) and pre-cooled 2 times volume of absolute ethanol to precipitate the obtained supernatant, centrifuge at 12000 rpm for 20 minutes to collect the precipitate; rinse with 70% ethanol twice Naturally dry at room t...

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Abstract

The invention belongs to the field of DNA markers in the field of aquaculture science and in particular relates to primers and a method for distinguishing Thai marble goby (oxyeleotrismarmoratus) fries from local marble goby (odontobutispotamophila) fries by adopting microsatellite markers. A pair of primer sequences is shown in SEQIDNo.8 and SEQIDNo.9. The method comprises the following steps of: carrying out PCR (polymerase chain reaction) amplification on the samples to be distinguished by utilizing primers; and then detecting the PCR products by using 1% of agarose gel electrophoresis and carrying out EB (ethidium bromide) staining, wherein the samples with 250bp stripes are Thai marble goby and the samples with 300bp stripes are local marble goby. The method has the following beneficial effects: the microsatellite markers obtained by the method can carry out stable amplification in oxyeleotrismarmoratus and odontobutispotamophila and have stable results and clear stripes; and the method is simple and convenient to operate and is low in cost and strong in practicability.

Description

Technical field [0001] The invention belongs to the application of DNA markup technology in the field of aquaculture in production, which involves the application of microbesarial marks to Thai bamboo shoots (cloud spotted tidam pond Oxyeleotris marmoatus ) And local bamboo shoots shellfish (riverside Shatang 鳢 Odontobutis Potamophila ) Fish fry is identified. Background technique [0002] Cloud spot tinson ( Oxyeleotris marmoatus ) The common name of the Thai bamboo shoot fish (Marble Goby), belongs to Pereifomes, Gobioidei, Eleotridae, and Shoutang ( Oxyeleotris ), Is the largest and fastest -growing fastest freshwater fish species in the fisher in the family of Tangyu.Yunba Tongtang is native to Southeast Asian countries such as Thailand, Vietnam, and Malaysia, and it is also one of the excellent varieties of exports to export foreign exchange.Because the fish is beautiful, delicious, tender, and rich in nutrition, it is welcomed by Chinese coastal cities, Hong Kong, Australia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 尹绍武朱晓平祝斐胡亚丽
Owner NANJING NORMAL UNIVERSITY
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