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Microsatellite marker applied to growth trait analysis of Macrobrachium nipponense and application thereof

A technology for microsatellite markers and growth traits, which is applied in the field of biological genetics and breeding of Macrobrachium prawns, can solve the problems of habitat disturbance, damage to the structure of population growth traits, and achieves the effect of stable PCR amplification results and assisted breeding.

Active Publication Date: 2017-01-25
HEBEI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, with the rapid advancement of industrialization, the habitat of Macrobrachium japonicus has been disturbed, destroyed and disorderly developed, resulting in the destruction of its population growth traits and structure, and the quality of germplasm resources has declined.
So far, there are few relevant studies on microsatellite markers of Macrobrachium japonicus in the prior art

Method used

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  • Microsatellite marker applied to growth trait analysis of Macrobrachium nipponense and application thereof
  • Microsatellite marker applied to growth trait analysis of Macrobrachium nipponense and application thereof
  • Microsatellite marker applied to growth trait analysis of Macrobrachium nipponense and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Measurement and Recording of Macrobrachium japonicus Population Growth Character Phenotype Values

[0032] 24 live samples of Macrobrachium japonicus were selected from Baiyangdian Lake (BYD) in Hebei, Hengshui Lake in Hebei (HSH), Weishan Lake in Shandong (WSH) and Hongze Lake in Jiangsu (HZH). Macrobrachium japonicus was weighed and measured, and the measurement indicators included weight, total length, body length, carapace length, carapace width, carapace height, abdomen length, abdomen width, abdomen height, tail fan length, There are 14 measurable traits including the width of the tail fan, the length of the forehead sword, the length of the second leg, and the length of the second leg. Schematic diagram of the measurement site for the morphological parameters of Macrobrachium japonicus figure 1 shown. The measurement is accurate to 0.02mm. in:

[0033] Body weight (BW): the wet weight of the whole body, use filter paper to absorb the water on the bo...

Embodiment 2

[0047] Example 2 Extraction of Total DNA from Macrobrachium japonicus Wild Population Muscle

[0048] The total muscle DNA was extracted using a marine animal genome extraction kit, and the specific steps were as follows:

[0049] (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.

[0050] (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 water droplets on the inner wall of the tube cap, and proceed to the next step.

[0051] (3) Add 200 μL buffer GB, mix thoroughly by inversion, place at 70°C for 10 minutes, the solution should become clear, and briefly centrifuge to remove water droplets on the inner wall of the tube cap.

[0052](4) Add 200 μL of absolute ethanol and mix thoroughly by inversion. At this time, f...

Embodiment 3

[0060] Example 3 Development of Microsatellite Marker Primers

[0061] (1) Macrobrachium japonicus was purchased from the Baiyangdian aquatic product market. The muscle tissue was taken under sterile conditions, and the total RNA of the muscle tissue was extracted, and sent to a biological company for transcriptome sequencing to obtain transcriptome data; the assembled length was obtained using MISA software. The unigenes over 1kb are identified for SSR. The identification criteria are: the minimum repeats of precise SSR markers containing two, three, four, five and six nucleotide types are 9, 6, 5, and 4 times respectively, using SSRHunter1.3 Perform SSR marker screening to ensure that the front and rear flanks of the sequence are of sufficient length for primer design. Use Primer Permier 6 to design primers based on the screened SSRs; the main parameters of the design are: the optimal length of the primer is 18-25 bp, the length of the PCR product fragment is 100-350 bp, the...

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Abstract

The invention discloses a microsatellite marker applied to growth trait analysis of Macrobrachium nipponense. The microsatellite marker comprises a microsatellite marker GL01 and a microsatellite marker GL02, wherein the microsatellite marker GL01 is correlated with a rostrum length trait; the microsatellite marker GL02 is correlated with a tail fan length trait, a second walking leg length trait and a second walking leg length knuckle trait; the nucleotide sequence of an upstream primer of the microsatellite marker GL01 is shown as SEQ ID NO:1; the nucleotide sequence of a downstream primer of the microsatellite marker GL01 is shown as SEQ ID NO:2; the nucleotide sequence of an upstream primer of the microsatellite marker GL02 is shown as SEQ ID NO:3; the nucleotide sequence of a downstream primer of the microsatellite marker GL02 is shown as SEQ ID NO:4. Meanwhile, the invention further discloses an application of the microsatellite marker to growth trait analysis of the Macrobrachium nipponense. The primers of the microsatellite marker have the characteristic of stable PCR (Polymerase Chain Reaction) amplification result, and the microsatellite markers can be applied to growth trait analysis of the Macrobrachium nipponense and can be used for assisting in molecular marker breeding of the Macrobrachium nipponense.

Description

technical field [0001] The invention relates to the field of biological genetic breeding of Macrobrachium prawn, in particular to a microsatellite marker for analyzing the growth traits of Macrobrachium japonicus and its application. 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 to 6) to form repeating units, such as (GA )n, (AC)n, (GAA)n (where n is the number of repetitions), etc., the number of repetitions is 10-50, and the same type of microsatellite DNA can be distributed in different positions of the whole genome, due to the difference in the number of repetitions or the degree of repetition The incompleteness of each locus forms a polymorphism. Most of the sequences at both ends of each type of microsatellite DNA are relatively conservative single-copy sequences, and a pair of specific primers can be designe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 康现江管越强武小斌穆淑梅
Owner HEBEI UNIVERSITY
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