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

A local bamboo shoot shell fish and bamboo shoot shell fish technology, applied in the field of aquaculture, can solve problems such as improper post-processing, and achieve low cost, clear strips, and strong practical effects

Inactive Publication Date: 2012-09-05
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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
  • 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 (Cherna chinensis population, Channa spp. population in rivers):

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

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

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

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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 andcarrying 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 themethod 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 marker technology in the field of aquaculture in production practice, and in particular relates to the application of microsatellite markers to Thai bamboo shoot shellfish (Cherna chinensis). Oxyeleotris marmoratus ) and local bamboo shoot shellfish (Channa chinensis Odontobutis potamophila ) fry for identification. Background technique [0002] Cloud-spotted sharp pond snakehead ( Oxyeleotris marmoratus ) common name Thai bamboo shoot shell fish (Marble goby), belongs to Pereiformes (Pereiformes), Gobioides (Gobioidei), pond snakehead family (Eleotridae), sharp pond snakeheads ( Oxyeleotris ), which is the largest and fastest-growing rare freshwater fish species in the Channaidae. The cloud-spotted snakehead is native to Southeast Asian countries such as Thailand, Vietnam, and Malaysia, and it is also one of the excellent varieties that are exported to these countries to earn foreign exchange. Bec...

Claims

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Application Information

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Patent Type & Authority Applications(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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