Sequence sequence repeats (SSR) molecular marker of thoracic window scirpus as well as primer combination and application thereof

A technology of molecular markers and primer combinations, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., can solve problems such as the development of molecular markers without firefly SSR

Pending Publication Date: 2022-07-15
武汉霆科生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] By studying the SSR molecular marker technology of the firefly, it can be used for paternity, genetic diversity, population genetics and germplasm identification and analysis of firefly offspring, and can also be used for sexual selection and evolution research of firefly insects, and Evaluation of artificial rearing, repopulation and protection of firefly firefly, however, so far, there is no relevant report on the development of firefly SSR molecular markers

Method used

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  • Sequence sequence repeats (SSR) molecular marker of thoracic window scirpus as well as primer combination and application thereof
  • Sequence sequence repeats (SSR) molecular marker of thoracic window scirpus as well as primer combination and application thereof
  • Sequence sequence repeats (SSR) molecular marker of thoracic window scirpus as well as primer combination and application thereof

Examples

Experimental program
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Embodiment 1

[0051] 1. Genome-wide SSR analysis and primer design of P.

[0052] The thoracic window firefly genome was de novo sequenced, and the assembled scaffold file was used as the input file. MISA was used to perform SSR scanning of the whole genome, and primer3 was used to design primers based on the 100 bp upstream and downstream flanking sequences of the detected SSR. Primer design conditions are as follows: optimal length 24bp; shortest length 20bp; longest length 28bp; optimal annealing temperature 63°C; minimum annealing temperature 60°C; maximum annealing temperature 65°C; °C. The number and types of SSR loci were analyzed, and then high-efficiency SSR primers that could be used for the identification of different male paternity were designed and developed. Considering the conservation and stability, the applicant mainly used microsatellites with 8 to 15 repeats of three bases, four bases and five bases. A total of 44 primer pairs were designed to identify SSR

[0053] 2. ...

Embodiment 2

[0068] The identification of paternity of fireflies on the chest window

[0069] Select females with a body length of 24.20±2.78mm, a body width of 7.27±0.59mm, and a body weight of 0.34±0.09g, as well as a body length of 17.73±1.35mm, a body width of 6.78±0.59mm, and a body weight of 0.07±0.02g One female firefly was selected to mate with 2 male fireflies (n=16), and the other group was designed to mate with one female firefly and 3 male fireflies (n=9), and lay eggs and hatch into larvae, respectively. Each female and corresponding mating males and all successfully hatched progeny larvae were collected for SSR analysis, using fluorescently labeled PCR primers to amplify multiple microsatellite polymorphic sites, and then using high-resolution gel electrophoresis. The amplified fragments were separated, and the length of the amplified fragments was determined by a fluorescence detection system (Licor 4300) to realize the typing of the microsatellite polymorphic sites. Using ...

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Abstract

The invention provides an SSR (Simple Sequence Repeat) molecular marker for a thoracic window, which is characterized in that the SSR molecular marker comprises one or more of 13 groups of SSR molecular markers, and the serial numbers of the 13 groups of SSR molecular markers are SSR308, SSR311, SSR313, SSR319, SSR320, SSR323, SSR324, SSR325, SSR405, SSR406, SSR501, SSR503 and SSR504 respectively; the nucleotide sequences of the 13 groups of SSR molecular marker sites are as shown in SEQ ID NO. 1 to 13. According to the SSR molecular marker disclosed by the invention, a technical system is established, so that the blank of the specific molecular marker of the scirpus juncoides is filled, the SSR molecular marker can be used for identifying and analyzing firefly offspring paternal right, genetic diversity, population genetics and germplasm, and a result can be used for sex selection and evolution research of a scirpus juncoides insect; and evaluating artificial feeding, rejuvenation and protection of the scirpus juncoides.

Description

technical field [0001] The invention relates to the technical field of DNA molecular markers, in particular to a thoracic window fluorescent SSR molecular marker, in particular to a thoracic window fluorescent SSR molecular marker and a primer combination and application thereof. Background technique [0002] Simple sequence repeats (SSRs, Simple Sequence Repeats), also known as microsatellites (Microsatellites), are sequences formed by tandem repeats of repeating units of 1-6 bp (Sharma et al. 2007). They are widely distributed in prokaryotic and eukaryotic genomes and can be found in both coding and non-coding regions of genes (Lietal. 2004). As a molecular marker, SSR has many advantages: it is widely distributed, uniform and sufficient in the whole genome, so it has high polymorphism; SSR markers are Mendelian co-dominant inheritance, which can distinguish homozygous and heterozygous genotypes; PCR amplification Growth requires only a small amount of DNA, and the qualit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2525/151C12Q2565/125
Inventor 付新华朱馨蕾刘全
Owner 武汉霆科生物科技有限公司
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