A kind of snp primer and detection method for genotyping different ginkgo biloba

A technique of genotyping and genotyping, applied in biochemical equipment and methods, microbiological determination/inspection, recombinant DNA technology, etc., can solve the problems of less research and unreported ginkgo research, and achieve simple operation methods and reduce The use of reagents and consumables, the effect of high sensitivity

Active Publication Date: 2019-10-15
NANJING FORESTRY UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the application of this technology in plants is mostly focused on crops, and there are relatively few studies on forest trees, and there is no report on the species of ginkgo.

Method used

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  • A kind of snp primer and detection method for genotyping different ginkgo biloba
  • A kind of snp primer and detection method for genotyping different ginkgo biloba
  • A kind of snp primer and detection method for genotyping different ginkgo biloba

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Experimental program
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Embodiment 1

[0035] 1. Preliminary screening of SNP primers for ginkgo genotyping

[0036] 1) Acquisition of transcriptome data

[0037] Ginkgo biloba total RNA extraction was performed using the RNeasy Plant Mini Kit (Qiagen, Hilde, Germany) according to the manufacturer's instructions. After extraction, RNA degradation and contamination were checked on a 1.0% agarose gel. use RNA purity was checked with a spectrophotometer (Implen, Westlake Village, CA, USA). use 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) The RNA AssayKit measures the concentration of RNA. RNA integrity was assessed using the RNA Nano 6000 Assay Kit in an Agilent Bioanalyzer 2100system (Agilent Technologies, Santa Clara, CA, USA).

[0038] A total of 3 μg RNA per sample was used as input material for RNA sample preparation. According to the manufacturer's instructions, use UltraTM RNA Library Prep Kit (NEB, USA) generated sequencing libraries and added index codes to the attribute sequences of ea...

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PUM

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Abstract

The invention discloses a SNP primer and a detection method for genotyping different ginkgoes. There are 22 pairs of primers in the present invention, and 11 pairs of SNP primer combinations can be used to construct the fingerprints of ancient and famous ginkgo trees. The SNP primers developed by the invention using ginkgo transcriptome data can genotype different ginkgo individuals, and the invention has simple operation method, high throughput and low cost. In addition, the 11 pairs of SNP primers developed by the present invention have constructed the fingerprints of ancient and famous ginkgo trees in different populations, and the verified 22 SNP sites can also be used for molecular marker-assisted breeding, genetic diversity research, and SNP-based association analysis , as well as identification and classification of germplasm resources.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a SNP primer and a detection method for genotyping different ginkgoes. Background technique [0002] Ginkgo biloba L., as the only extant species of gymnosperm Ginkgopsida, is a typical Mesozoic relict species and is known as a "living fossil". Ginkgo biloba is a large deciduous tree with tall trunks and dioecious plants. The bark is light green when young, taupe when old, deeply longitudinally fissured, and rough; the branches are luxuriant, divided into long and short branches. Seeds are drupe-like and smelly; the middle testa is bony, white, with 2-3 ribs; the inner testa is membranous light brown, with 2 cotyledons; the flowering period is April-May, and the seeds mature in September-October. Ginkgo is an all-round tree species that integrates timber, fruit, leaf, landscaping, and ornamental. Ginkgo biloba contains a large amount of ginkgo flavonoids and ginkgolid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 徐立安吴雅琼王建文胥猛周琦
Owner NANJING FORESTRY UNIV
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