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Anthurium ssr primer pair and kit developed based on transcriptome sequencing

A technology of transcriptome sequencing and primer pairs, applied in the field of SSR primers, can solve problems affecting accuracy and precision

Active Publication Date: 2016-08-17
ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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AI Technical Summary

Problems solved by technology

The number of molecular markers affects the accuracy of genetic diversity analysis of germplasm resources and the accuracy of genetic map construction. The number of existing Anthurium SSR molecular markers is far from meeting the needs of Anthurium molecular biology research.

Method used

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  • Anthurium ssr primer pair and kit developed based on transcriptome sequencing
  • Anthurium ssr primer pair and kit developed based on transcriptome sequencing
  • Anthurium ssr primer pair and kit developed based on transcriptome sequencing

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Embodiment Construction

[0036] Anthurium SSR primer pairs developed based on transcriptome sequencing were obtained through the processes of Anthurium anthurium transcriptome sequencing, SSR primer development, DNA extraction, SSR primer screening and variety identification ( figure 1 ).

[0037] In the present invention, the specific practice of developing Anthurium SSR primer pair with the EST sequence obtained by transcriptome sequencing is:

[0038] 1. Anthurium Transcriptome Sequencing

[0039] After extraction of total RNA from Anthurium (Alabama) leaves by Trizol method, eukaryotic mRNA was enriched with magnetic beads with Oligo(dT). Fragmentation buffer was added to break the mRNA into short fragments, and the mRNA was used as a template to synthesize the first cDNA strand with random hexamers, and then add buffer, dNTPs, RNase H and DNA polymerase I to synthesize the second cDNA After purification by QiaQuick PCR kit and elution with EB buffer, end repair was performed, poly(A) was added...

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Abstract

The invention relates to anthurium SSR (Simple Sequence Repeat) primer pairs and a kit based on transcriptome sequencing development. The invention provides sequences of 22 anthurium SSR primer pairs and a kit for identifying genetic relationship and hereditary properties of anthurium. The 22 primer pairs have polymorphism in 50 anthurium species, and match the hereditary diversity analysis on the 50 anthurium species with common knowledge of plant classification. The 22 primer pairs can be directly applied to more anthurium plant species to perform hereditary diversity analysis on germ plasm resources.

Description

technical field [0001] The present invention relates to SSR primers, in particular to a set of SSR primer pairs and kits developed based on Anthurium transcriptome sequencing, and belongs to the field of biotechnology. Background technique [0002] Anthurium is a perennial evergreen herb of the genus Anthurium in the Araceae family. Its flower shape is beautiful, its flowering period is long, and its ornamental value is high. It ranks second only to orchids in the global tropical flower trade. Anthurium has been introduced in my country since the 1990s, and it has become one of the main flowers in my country. There are more and more germplasm types, which are mainly distinguished from the aspect of morphological characteristics. The phenomenon of homozygosity and heterogeneity of the same name is common. , can not reflect the real kinship and genetic characteristics, which will affect the collection, preservation and utilization of varieties, and bring difficulties to the cul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 葛亚英田丹青金亮沈晓岚潘晓韵刘晓静
Owner ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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