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Multiple genotyping kit for detecting genetic quality of zebra fish

A technology for detection kits and quality detection methods, applied in the field of multiple typing scheme kits

Inactive Publication Date: 2012-02-08
SHANGHAI RES CENT FOR MODEL ORGANISMS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] As a direct carrier of genetic information, DNA molecules are not affected by the environment, biological developmental stages, and organ tissues, so they can more accurately reflect the phylogenetic relationship between various species. However, the conventional PCR method requires a PCR reaction to amplify

Method used

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  • Multiple genotyping kit for detecting genetic quality of zebra fish
  • Multiple genotyping kit for detecting genetic quality of zebra fish
  • Multiple genotyping kit for detecting genetic quality of zebra fish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The multiple PCR amplification of embodiment 1 sample DNA

[0021] (1) Amplification system (5μl):

[0022] Primers (0.5μM / pair)×n 0.05μM each

[0023] 10×Buffer 1×

[0024] dNTP 0.2mM

[0025] Mg 2+ 3.0mM

[0026] Taq polymerase 0.25U

[0027] Add water to 5 μl

[0028] (2) Amplification program:

[0029] 95℃15min

[0030] 94℃30s→50~65℃90s→72℃60s(35cycles)

[0031] 72℃10min→4℃forever

Embodiment 2

[0032] Ligase Detection Reaction (LDR) of Example 2 Sample DNA

[0033] (1) The PCR product amplified by PCR primers is used for LDR signal detection

[0034] (2) LDR system (5μl):

[0035] Site-specific probes: 0.1 μM each; Universal fluorescent probes: 0.1 μM;

[0036] Butter 1×

[0037] Template 2μl

[0038] Taq 2U

[0039] h 2 O supplemented to 5 μl

[0040] (3) LDR reaction procedure:

[0041] 95℃2min

[0042] 94℃30s→50℃2min(30cycles)

Embodiment 3

[0043] Example three probe specificity assessment

[0044] (1) SNP specificity assessment

[0045] The selected 13 SNP loci were used to test the samples of known foreign zebrafish strains for SNP specificity evaluation. The foreign zebrafish strains are relatively pure and can be used to detect the genetic quality of domestic zebrafish.

[0046]

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Abstract

The invention discloses a multiple genotyping kit for detecting the genetic quality of zebra fish. Lines of the zebra fish are diversified; and experimental zebra fish is extremely important for the research on the fields of development biology, genetics, oncology, pharmacology, toxicology, environment friendliness and the like. Particularly, the germplasm degeneration and miscegenation of the zebra fish can seriously affect the scientificity of experimental results, so the kit has a great scientific significance for detecting the genetic quality of the zebra fish. A multiple genotyping scheme kit for a method for detecting the genetic state of the zebra fish is established by taking four lines of the zebra fish, namely TU, TL, AB and WIK as research objects, designing primers and probes for known single nucleotide polymorphism (SNP) sites and utilizing an SNP site primer polymerase chain reaction (PCR) combined ligase detection reaction (LDR) technology.

Description

technical field [0001] The invention relates to the field of zebrafish genetic quality detection methods, in particular to selecting 12 SNP sites for 4 commonly used strains of zebrafish, designing 12 pairs of primers and specific probes, and using SNP site primer PCR combined with ligase detection reaction (ligase detection reaction, LDR) multiple typing protocol kit for detecting the genetic quality of zebrafish. Background technique [0002] At present, zebrafish is extremely important to the research of developmental biology, genetics, oncology, pharmacology, toxicology and environmental protection. At present, in addition to the quality of microbiology and parasitology, domestic zebrafish also has weaknesses in genetic quality. Several relatively mature zebrafish strains have already been formed in the world, and they are used as a new breeding method for exploration. In the introduction and conservation of zebrafish in many domestic units, there are not only blind in...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 胡建华赵莹濮俊毅沈志敏刘雄伟
Owner SHANGHAI RES CENT FOR MODEL ORGANISMS
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