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Method for distinguishing black snapper and two hybrid filial generations of black snapper by using ISSR molecular marker

A technology of molecular markers and black snapper, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as no detection methods, and achieve high accuracy, good versatility, The effect of easy operation

Pending Publication Date: 2022-05-13
INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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  • Abstract
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  • Claims
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Problems solved by technology

So far, there is no report on the relevant detection methods that can simultaneously identify the three germplasms of black sea bream, BF1 and HF1

Method used

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  • Method for distinguishing black snapper and two hybrid filial generations of black snapper by using ISSR molecular marker
  • Method for distinguishing black snapper and two hybrid filial generations of black snapper by using ISSR molecular marker

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Embodiment 1

[0023] 1) Three healthy individuals were randomly selected from each of black sea bream, hybrid offspring, and backcross offspring. Cut a mung bean-sized fish (or other body tissue), cut it into pieces with scissors and put it into a 1.5mL centrifuge tube; use the animal cell / tissue genomic DNA extraction kit or the phenol-chloroform method to extract the total DNA of each individual , stored at 4°C.

[0024] 2) ISSR primers: UBC847, UBC846 in the ninth set of primers ISSR (http: / / www.biotech.ubc.ca / services / naps / primers / Primers.pdf) of the University of Columbia (UBC), UBC846, primer (UBC847 Sequence: CAC ACA CAC ACA CAC ARC, UBC846-CAC ACA CAC ACA CAC ART) was synthesized by Shanghai Sangong.

[0025] 3) The total volume of the PCR reaction for three types of ISSR primers including black sea bream, hybrid progeny and backcross progeny is 25 μl, including: 1 μl primer (10 pmol / L), DNA template about 1-1.5 μl (15-20ng ), Buffer 2.5μl, dNTP 0.5μl, Mg 2+ 1.5 μl, Taq polymeras...

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Abstract

The invention relates to an ISSR (inter-simple sequence repeat) molecular marker method for distinguishing black sea bream and two hybrid filial generations of the black sea bream, which comprises the following steps: carrying out PCR (polymerase chain reaction) amplification on individuals of the black sea bream and two new hybrid lines by using two ISSR molecular markers, and distinguishing the individuals of three kinds of black sea bream from the black sea bream, the hybrid filial generations and the backcross filial generations according to electrophoretic band detection on PCR products. The method disclosed by the invention is simple and convenient to operate, low in cost, high in accuracy and good in universality, can quickly and effectively identify a new hybrid strain obtained based on different parent combinations of the black sea bream, and provides technical help for cultivation and research of a new black sea bream variety and identification and research of a new hybrid germplasm.

Description

technical field [0001] The invention belongs to the field of identifying black sea bream, in particular to a method for distinguishing black sea bream and its two hybrid progeny by using ISSR molecular markers. Background technique [0002] Black sea bream (Acanthopagrus schlegelii, AS) is a valuable economic fish in the family Sparidae and the genus Acanthopagrus. Black sea bream has the characteristics of broad temperature, strong disease resistance and delicious taste, and is an important fish cultured in my country's coastal areas. The slow growth rate of black sea bream restricts the development of its aquaculture industry; and the red sea bream (Pagrus major), which is also a member of the sea bream family, is a species of red sea bream (Pagrus), which grows fast and has a beautiful body color. Breeding objects. In order to select and breed new varieties of excellent marine aquaculture, we have obtained new germplasm with the advantages of target traits such as fast ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/124C12Q2565/125
Inventor 陈淑吟张志勇吉红九贾超峰孟乾祝斐徐大凤张志伟尹绍武张虎高波孙瑞健赵永超肖李霞
Owner INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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