Microsatellite markers related to growth character of portunus trituberculatus and primers and application thereof

A technology of microsatellite marking and swimming crabs three warts, which is applied in the field of microsatellite marking, can solve the problems of declining breeding area, worsening water quality, reducing the number of high-quality parents, etc., and achieves the effects of stable PCR amplification results and assisted breeding.

Active Publication Date: 2019-11-22
HEBEI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, industrial pollution in coastal areas has led to poor water quality, which has destroyed the living environment of Portunus trituberculatus, sharply reduced the number of high-quality parents, and greatly reduced the breeding area.

Method used

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  • Microsatellite markers related to growth character of portunus trituberculatus and primers and application thereof
  • Microsatellite markers related to growth character of portunus trituberculatus and primers and application thereof
  • Microsatellite markers related to growth character of portunus trituberculatus and primers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 3

[0024] Example 1 The source of Portunus trituberculatus population samples and the measurement of growth trait phenotype values

[0025] Three species from Liaoning Dalian (DL), Liaoning Huludao (HLD), Hebei Qinhuangdao (QHD), Hebei Huanghua cultured population (HHYZ), Hebei Huanghua wild population (HHYS), Shandong Dongying (DY) and Shandong Penglai (PL) were selected. 60 live and fresh samples of Portunus verruca. Each sample of Portunus trituberculatus was weighed and measured, and the measurement indicators included carapace width (BJK), carapace width of the eighth pair of spines (BBK), and inner carapace width (NBK) , carapace base width (BDK), carapace length (BJC), forehead width (QEK), forehead middle spine distance (QZJ), forehead side spine distance (QCJ), plastron width (FJK), belly width (FBK) , the length of the 3rd-step leg length (3CC), the 4th-step toe-joint length (4ZC), the 4th-step toe-joint width (4ZK), the length of the movable phalanx (KZC), and the leng...

Embodiment 2 3

[0026] Example 2 Extraction of Portunus trituberculatus sample muscle total DNA

[0027] The total muscle DNA was extracted using a marine animal genome extraction kit, and the specific steps were as follows:

[0028] (1) Cut no more than 30 mg of tissue material, put it into a centrifuge tube filled with 200 μL GA buffer, and vortex for 15 seconds.

[0029] (2) Add 20 μL of Proteinase K (20mg / mL) solution, vortex to mix, and briefly centrifuge to remove water droplets on the inner wall of the tube cap. Place at 56°C until the tissue is completely dissolved, centrifuge briefly to remove the water droplets in the tube cap, and then proceed to the next step.

[0030] (3) Add 200 μL buffer GB, mix thoroughly by inversion, place at 70°C for 10 min, the solution should become clear, and briefly centrifuge to remove water droplets on the inner wall of the tube cap.

[0031] (4) Add 200 μL of absolute ethanol and mix thoroughly by inversion. At this time, flocculent sediment may ap...

Embodiment 3

[0039] The development of embodiment 3 microsatellite primers

[0040] (1) Portunus trituberculatus was purchased from the original species of Portunus trituberculatus in Huanghua, its muscle tissue was taken under aseptic conditions, total RNA was extracted from the muscle tissue, and sent to a biological company for transcriptome sequencing to obtain transcriptome data; the assembly was performed using MISA software The obtained unigenes with a length of more than 1 kb were identified for SSR, and the identification criteria were: the minimum repeat numbers of precise SSR markers containing two, three, four, five and six nucleotide types were 9, 6, 5, and 4 times, respectively , using SSR Hunter 1.3 to screen SSR markers to ensure that the front and rear flanks of the sequence have sufficient length for designing primers. Use Primer Permier 6 to design primers based on the screened SSRs; the main parameters of the design are set as follows: the primer length is 18-25bp as th...

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Abstract

The invention provides microsatellite markers related to a growth character of portunus trituberculatus, and primers and an application thereof. The microsatellite markers include microsatellite markers ZL03, ZL05, ZL06, ZL08 and NC07. The invention also provides the application of the microsatellite markers or the primers thereof in analysis of the growth character of portunus trituberculatus andmolecular breeding of portunus trituberculatus. The primers of the microsatellite markers provided by the invention have the characteristics of stable PCR amplification results, and the microsatellite markers can be used for analysis of the growth characterof portunus trituberculatus, and can assist in breeding of the molecular markers of portunus trituberculatus.

Description

technical field [0001] The invention relates to the field of biological genetic breeding of Portunus trituberculatus, in particular to a microsatellite marker related to growth traits of Portunus trituberculatus, primers and application thereof. Background technique [0002] SSR (Simple sequence repeat) is a simple sequence repeat, also known as microsatellite DNA (microsatellite DNA). In eukaryotic genomes, SSR consists of only a few nucleotides (1-6) to form repeating units, such as (( CT)n, (AC)n, (ACT)n (where n is the number of repetitions), etc., the number of repetitions is 10-20, and the same type of microsatellite DNA can be distributed in different positions throughout the genome, due to the difference in the number of repetitions, or The degree of incompleteness forms the polymorphism of each locus. The sequences at both ends of each type of microsatellite DNA are mostly relatively conservative single-copy sequences. A pair of specific primers can be designed acco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 康现江段保华管越强穆淑梅姬新东李泽健李书琴李彦芹
Owner HEBEI UNIVERSITY
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