Method for identifying carassius auratus ploidy by using microsatellite marker and application thereof

A microsatellite marker and microsatellite technology, applied in the field of molecular biology, can solve problems such as time-consuming and laborious, and achieve the effects of strong operability, high sensitivity and rapid identification

Active Publication Date: 2018-09-07
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

[0003] The identification of crucian carp polyploidy can be done by chromosome preparation or flow cytometry. Although the result is accurate, it is time-consu

Method used

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  • Method for identifying carassius auratus ploidy by using microsatellite marker and application thereof
  • Method for identifying carassius auratus ploidy by using microsatellite marker and application thereof
  • Method for identifying carassius auratus ploidy by using microsatellite marker and application thereof

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Experimental program
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Embodiment 1

[0028] Test object: wild crucian carp from Taihu Lake was selected, one-on-one breeding, 12 full-sib families were constructed, and 5 fish were randomly selected from the offspring of each family, a total of 60 fish were used for polyploidy identification.

[0029] A method for identifying crucian carp ploidy by microsatellite markers, said method comprising the following steps:

[0030] (1) Develop microsatellite sequences, design and screen primers. The 11 pairs of microsatellite primers selected are: Caraur1, SEQ ID NO.1~2; Caraur 2, SEQ ID NO.3~4; Caraur3, SEQ ID NO.5~ 6; Caraur4, SEQ ID NO.7~8; Caraur5, SEQ ID NO.9~10; Caraur6, SEQ ID NO.11~12; Caraur7, SEQ ID NO.13~14; ​​Caraur8, SEQ ID NO.15~ 16; Caraur9, SEQ ID NO.17-18; Caraur10, SEQ ID NO.19-20; Caraur11, SEQ ID NO.21-22;

[0031] The above primers are fluorescently labeled, specifically: the fluorescent dye markers of Caraur1, Caraur4, Caraur7, Caraur8, and Caraur11 are FAM; the fluorescent dye markers of Caraur 2,...

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Abstract

The invention provides a method for identifying carassius auratus ploidy by using a microsatellite marker and application thereof. The method comprises the following steps that (1) a microsatellite sequence is developed, and a primer is designed and screened; (2) a carassius auratu tail fin ray is collected, and genome DNA is extracted; (3) the fin ray genome DNA is taken as a template to performPCR amplification; (4) capillary electrophoresis detection is performed on a PCR amplified product obtained in step (3) by using an ABI3730 sequenator; (5) a peak value graph of the ABI3730 sequenatoris analyzed. Compared with the prior art, the method for identifying the carassius auratus ploidy by using the microsatellite marker and the application thereof have the advantages that (1) the microsatellite marker has the advantages of high specificity, high sensitivity and good amplification efficiency; (2) compared with a conventional karyotype and streaming cell method, the method has the advantages that the operability is strong, the cost is reduced, and the time is shortened; and (3) the method can accurately and quickly identify the carassius auratus ploidy on the premise of not damaging a carassius auratu body.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a method for identifying crucian carp germplasm using a molecular marker method, in particular to a method for identifying crucian carp ploidy by microsatellite markers and an application thereof. Background technique [0002] Crucian carp belongs to Cyprinidae, Cyprinus subfamily, and the genus Crucian carp in classification. It is widely distributed in China, Japan, and the Korean Peninsula. After transplantation, it was introduced and domesticated to India, North America, and other parts of the world. It has strong adaptability and is currently distributed all over the world. There are two subspecies of crucian carp: crucian carp (Carassius auratus) and gibel crucian carp (Carassius auratus gibelio), as well as many varieties such as red crucian carp, white crucian carp, and goldfish. They have different numbers of chromosomes and different ploidy. At present, it is be...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 唐永凯王美垚李建林刘凯李红霞于凡俞菊华
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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