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Hippocampus erectus microsatellite markers and screening method thereof

A technology of microsatellite markers and lined hippocampus, which is applied in the field of microsatellite markers, can solve the problems of lack of molecular markers of lined hippocampus, and achieve the effects of enriching polymorphism, saving manpower and cost, and efficient construction methods

Active Publication Date: 2015-12-23
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem solved by the present invention is to provide a microsatellite marker and its screening method for the lack of existing linear hippocampal molecular markers

Method used

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  • Hippocampus erectus microsatellite markers and screening method thereof
  • Hippocampus erectus microsatellite markers and screening method thereof
  • Hippocampus erectus microsatellite markers and screening method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Acquisition and verification of microsatellite loci and specific primers in the striated hippocampus:

[0035] Collection of lined hippocampus samples and RNA extraction:

[0036] Select 30 healthy seahorses with well-developed gonads from a seahorse breeding company, cut off their dorsal fin rays, and immediately extract total RNA. The specific steps are as follows: (1) Put about 50 mg of the fins of the lined seahorses into a pre-cooled mortar , use liquid nitrogen to quickly grind into powder, put it into a 2mL centrifuge tube, and add 1mL TRIzol; (2) add 200μL chloroform to the centrifuge tube, shake vigorously for 15s, and let it stand at room temperature for 3min until the solution is separated; (3) 4°C, Centrifuge at 12,000r / min for 15min, absorb the supernatant and put it into a new centrifuge tube; (4) add isopropanol equal to the volume of the supernatant, shake gently to mix, and place at room temperature for 5-10min; (5) 4°C, 12,000 Centrifuge at...

Embodiment 2

[0049] Embodiment 2. The family identification test of striae hippocampus:

[0050] Establishment of lined hippocampus pedigree and sample collection:

[0051] Select 30 pairs of healthy seahorses with well-developed gonads from a seahorse breeding company, put 1 female and 1 male seahorse in each bucket, and induce aphrodisiac, oviposition, pregnancy and young under artificial ecological conditions. After the male seahorses hatch young seahorses, immediately cut the dorsal fin rays of the female and male seahorses in the paired group, and collect the newly born seahorses, 10-15 for each family, and store them in 100% alcohol as a family identified material.

[0052] DNA extraction of lined hippocampus parents and offspring:

[0053] The ammonium acetate / isoamyl alcohol method was used to prepare the DNA of the lined hippocampus, and the specific steps were as follows: (1) put about 50 mg of the fins of the lined parent and the whole juvenile hippocampus into a 2 mL centrifu...

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Abstract

The invention belongs to the technical field of fish breeding, and discloses Hippocampus erectus microsatellite markers and a screening method thereof. The Hippocampus erectus microsatellite markers are disclosed as SEQ.ID.NO:1-9. The screening method comprises the following steps: 1) carrying out transcriptome sequencing screening to obtain Hippocampus erectus Unigenes; 2) detecting microsatellite sites in the Unigenes sequence; 3) designing specific primers according to sequences on both ends of the microsatellite site; and 4) carrying out PCR (polymerase chain reaction) amplification and detecting the polymorphism, thereby obtaining the 9 microsatellite markers. The invention discloses the Hippocampus-erectus-related microsatellite markers and specific primers thereof for the first time. The Hippocampus erectus microsatellite markers can be used for researching Hippocampus erectus population genetic structure, genetic breeding, identification in disputed paternity and the like.

Description

Technical field: [0001] The invention belongs to the technical field of microsatellite markers, in particular to a linear hippocampal microsatellite marker and a screening method thereof. Background technique: [0002] Hippocampus, belonging to the family Syngthidae and Hippocampus, is a rare marine Chinese herbal medicine known as "Southern Ginseng" in my country. , At the same time, it also has high ornamental value and the function of collecting decorations. In the past few decades, due to factors such as long-term overfishing, environmental pollution and climate change, seahorse germplasm resources have been threatened by serious degradation, and some seahorse species have even reached the edge of extinction. In order to effectively protect the germplasm resources of seahorses, 38 species of seahorses were included in the 2012 Red List of Threatened Animals by the International Union for Conservation of Nature (IUCN). Therefore, it is very important to study the genetic...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 罗伟林强屈宏越王信
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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