Anthurium SSR (Simple Sequence Repeat) primer pairs and kit based on transcriptome sequencing development

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: 2014-12-10
ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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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 (Simple Sequence Repeat) primer pairs and kit based on transcriptome sequencing development
  • Anthurium SSR (Simple Sequence Repeat) primer pairs and kit based on transcriptome sequencing development
  • Anthurium SSR (Simple Sequence Repeat) primer pairs and kit based on transcriptome sequencing development

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Embodiment Construction

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

[0037] In the present invention, the specific method of developing anthurium SSR primer pairs with the EST sequence obtained by transcriptome sequencing is:

[0038] 1. Anthurium transcriptome sequencing

[0039] After extracting total RNA from leaves of Anthurium (Alabama) by Trizol method, the eukaryotic mRNA was enriched with Oligo(dT) magnetic beads. Add fragmentation buffer to break mRNA into short fragments, use mRNA as a template, use six base random primers (random hexamers) to synthesize the first cDNA strand, then add buffer, dNTPs, RNase H and DNA polymerase I to synthesize the second strand The cDNA chain is purified by QiaQuick PCR kit and eluted with EB buffer, then repaired at the end, added poly(A) and c...

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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 invention relates to SSR primers, specifically a set of SSR primer pairs and a kit developed based on Anthurium transcriptome sequencing, belonging to the field of biotechnology. Background technique [0002] Anthurium is a perennial evergreen herb of the Araceae family Anthurium. It has beautiful flower shape, long flowering period and high ornamental value. 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 produced in my country. There are more and more types of germplasm, which are mainly distinguished from morphological characteristics. The phenomenon of the same species with different names and the same name with different species is common. , can not reflect the true genetic relationship and genetic characteristics, which will affect the collection, preservation and utilization of varieties, and bring difficulties to the cu...

Claims

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Application Information

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Patent Type & Authority Applications(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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