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Method for rapidly determining genome size of sturgeons by flow cytometry

A flow cytometry and rapid determination technology, which is applied in the field of determination of the genome size of sturgeon species, can solve the problems of complicated processing technology, infection risk of incisions, increased feeding costs, etc., and achieve the effect of rapid determination

Inactive Publication Date: 2018-11-16
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] ①, greatly increased the cost of feeding
[0006] ②, increasing the cost of gender identification
[0007] ③. Identify the risk of infection in the incision made
Yin Hongbin and others used the tail fin of fish as the sample to be tested, and the sample processing process was relatively complicated.

Method used

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  • Method for rapidly determining genome size of sturgeons by flow cytometry
  • Method for rapidly determining genome size of sturgeons by flow cytometry
  • Method for rapidly determining genome size of sturgeons by flow cytometry

Examples

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Embodiment 1

[0047] Embodiment 1, the method for rapidly measuring the genome size of sturgeon by flow cytometry, carries out the following steps successively:

[0048] 1. Grass carp, a carp with known genome size, was selected as a standard control. The genome size of grass carp is about 1.07 Gb. Steps 1) to 7) of the following operations are performed on the grass carp and the sturgeons to be tested (female Acipenser dauribus, male Acipenser dauribus, female Acipenser schneidere, male Acipenser schneidere, female Russian sturgeon and male Russian sturgeon):

[0049] 1) Take blood from the tail vein, take 0.5ml of blood, and store it in EDTA anticoagulant tubes.

[0050] 2) Take fresh blood or anticoagulated blood and centrifuge at 1500rpm for 5min. Wash twice with PBS buffer (remove the washing solution after each wash, the same below), add 0.5ml PBS buffer to resuspend red blood cells; obtain red blood cell suspension I.

[0051] 3) Take 50 μl of the erythrocyte suspension I obtained ...

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Abstract

The invention discloses a method for rapidly determining the genome size sturgeons of by flow cytometry. The method comprises the following steps: 1, selecting fish with the genome having a known sizeas a standard control, respectively performing caudal vein blood connection, blood centrifuging, PBS buffer solution washing, 2% paraformaldehyde solution addition, permeabilizing agent addition andstaining on the standard control and a sturgeon to be detected, and detecting the standard control and the sturgeon to be detected by flow cytometry to obtain the fluorescence intensity peak P2 of thestandard control and the fluorescence intensity peak P1 of the sturgeon to be detected; and 2, obtaining the genome size of the sturgeon to be detected according to N1= N2 * P1 / P2, wherein N1 represents the genome size of the sturgeon to be detected, and N2 represents the genome size of the standard control. The method can be used to preliminary determine the genome size of several major speciesof sturgeons bred in China before large-scale genome sequencing.

Description

technical field [0001] The invention belongs to the field of fish genomics, and specifically refers to a method for measuring the genome size of several major sturgeon species in my country by using flow cytometry. Background technique [0002] Sturgeon (Acipenser sturio Linnaeus) belongs to the order Acipenseridae and Acipenseridae, belonging to the subclass Actinopterygia Acipenseridae, and includes 26 species in 6 genera. Sturgeon is one of the oldest species of fish in the world, with a history of more than 200 million years, known as the "living fossil in water". Sturgeons are mainly distributed in the North American continent and the Eurasian continent in the northern hemisphere. There are 8 species in my country, 3 species are distributed in Xinjiang (Siberian sturgeon, naked-bellied sturgeon, and small sturgeon), and 2 species are distributed in Heilongjiang (Huso dabryi, Sturgeon's sturgeon). ), 2 species in the Yangtze River (white sturgeon, Dabry's sturgeon), and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
CPCG01N15/1434G01N2015/1006G01N2015/1486G01N2015/1493G01N15/01
Inventor 梅祎芸郑荣泉王志刚
Owner ZHEJIANG NORMAL UNIVERSITY
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