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Microsatellite fluorescence multi-PCR method for testing paternity of culter alburnus basilewsky

A technology for paternity identification and tuna, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effects of high experimental efficiency, saving DNA samples, and simple detection methods

Active Publication Date: 2018-10-02
ZHEJIANG INST OF FRESH WATER FISHERIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology allows researchers to detect specific areas on chromosomal DNA called microspots (DNA fragments) that may indicate different traits or diseases like cancer. It simplifies this process by saving only necessary materials while still being able to accurately identify these regions within an individual's genome.

Problems solved by technology

This patented describes various techniques related to studying microscopic organisms such as bacteria or plants called cullstrahols during their life cycle from seafoods through turtles on land. These methods require many steps like extracting nucleic acid material from them, preparation of reagents, analysis by sequencing, etc., resulting in significant expenses associated with each step. Additionally, these conventional approaches involve analyzing numerous small sections of DNA obtained from several genera, making it difficult to identify specific sequences within those segments without consuming too much data.

Method used

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  • Microsatellite fluorescence multi-PCR method for testing paternity of culter alburnus basilewsky
  • Microsatellite fluorescence multi-PCR method for testing paternity of culter alburnus basilewsky

Examples

Experimental program
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Effect test

Embodiment 1

[0026] (1) Select the 3-year-old C. chinensis with mature gonads for artificial induction, and establish 12 full-sibling single-pair mating families; collect all parent individuals (24 in total) and some offspring individuals (20 for each family) Splashes, 480 in total);

[0027] (2) Take the fin ray of the Culturus chinensis individual, use the phenol-chloroform method to extract the genomic DNA, measure the concentration of the obtained DNA, and then dilute it to about 50 ng / μl;

[0028] (3) Screening of polymorphic microsatellite sequences

[0029] According to the NCBI sequence recorded in the NCBI, the Primer Premier 6.0 software was used to design the primer sequence of the Cichlid chinensis and the existing literature records, and to ensure that there is a large difference in the length of the target fragment (100-500bp), the primer sequence was sent to the biological company for synthesis. Using the genomic DNA obtained in step (1) as a template, the synthesized prime...

Embodiment 2

[0041] (1) Collect 32 cultured individuals in the Huzhou farm;

[0042] (2) Use the phenol-chloroform method to extract the DNA of each sample, and then dilute the obtained DNA to about 50 ng / μl after measuring its concentration;

[0043] (3) Synthesize primers according to the 12 kinds of primer sequences screened above and the modification method, the method is the same as that in Example 1;

[0044] (4) Perform multiple PCR amplification using the aforementioned three primer combinations, and the multiple PCR amplification system and amplification conditions are the same as in Example 1;

[0045] (5) After the PCR amplification product is properly diluted, it is mixed with a molecular weight internal standard and analyzed by capillary electrophoresis on an ABI 3130 sequencer;

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Abstract

The invention relates to a microsatellite fluorescence multi-PCR method for testing the paternity of culter alburnus basilewsky. The method comprises the following steps: firstly, 16 pairs of SSR primers are screened out, 12 pairs of SSR primers are chosen through optimal combination to form 3 multi-PCR groups; 4 pairs of SSR specific primer pairs belonging to the same group but having different concentrations are then added into a PCR reaction system, different reaction systems are used to amplify different areas of one same DNA template to form a plurality of target fragments; the target fragments are sent to a biological company for typing on a sequencer after the sampling inspection through electrophoresis, a genotype on a corresponding lotus of a sample is acquired; and finally the genotype is used to analyze the parentage of the parent and the progeny. A related microsatellite marker amplification system saves the DNA sample, is high in experiment efficiency, and can be popularized on the paternity of culter alburnus basilewsky as well as evaluation of genetic variation of population and hereditary constitution.

Description

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Claims

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Application Information

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Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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