Li SSR marker primer pair developed based on transcriptome sequence and its application

A technology of transcriptome sequence and labeling primers, which is applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., to achieve the effect of increasing original data and improving the efficiency of development

CN105238781BActive Publication Date: 2018-02-16POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2018-02-16

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Abstract

The invention belongs to the field of molecular marker technology development and application, and specifically provides the development of plum SSR marker primer pairs based on transcriptome sequences and its application in the research of plum germplasm resources. The primer pair is developed based on the transcriptome sequence. On the basis of transcriptome sequencing, a large amount of sequence information is processed in batches, SSR site search and SSR primer design are performed, and SSR markers can be developed in large quantities, which greatly improves the development efficiency. It overcomes the problems of complex steps, heavy workload, and high development cost in developing SSR markers in the traditional way. Finally, SSR primer screening and diversity analysis were carried out to verify the effectiveness of SSR primers. The invention lays a foundation for the development of plum SSR marker primers and the use of SSR molecular markers for plum germplasm resource diversity, variety identification and genetic relationship research.
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Description

Technical field

[0001] The invention belongs to the field of molecular marker technology development and application, and specifically relates to a plum SSR marker primer pair developed based on a transcriptome sequence and a method for SSR primer development. Background technique

[0002] Prunus is a plant of the genus Prunus in the Rosaceae family. It is widely distributed throughout the world and is also one of the important traditional fruit trees in my country. Plum fruits are rich in bioactive substances such as carotenoids, vitamins and anthocyanins, which have the effect of preventing many diseases and are deeply loved by consumers. The plum germplasm resources in my country are extremely rich. There are many plum varieties, the naming is unscientific, and the phenomenon of the same name or the same thing with different names is very serious, which seriously hinders the development and utilization of plum germplasm resources. Therefore, it is very important to collect, p...

Examples

Embodiment 1

[0058] The plum SSR primer pair developed based on transcriptome sequencing is obtained through plum transcriptome sequencing, SSR site identification, SSR primer design, DNA extraction, SSR primer screening, and genetic diversity analysis.

[0059] In the present invention, the specific method of the plum SSR primer pair developed based on the transcriptome sequence is:

[0060] 1) Acquisition of transcriptome data

[0061] EZNA Plant RNA Kit (Omega Bio-tek) was used for the extraction of total RNA from hibiscus plum fruit. The RNA concentration was determined by Qubit 2.0 Fluorometer (Invitrogen, Life Technologies, CA, USA), and the quality of RNA was evaluated by Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The RNA from the fruit samples of the same stage of three different trees was mixed in equal amounts and used for sequencing library construction.

[0062] The EZNA Plant RNA Kit (Omega Bio-tek) was used to extract the total RNA from the hibiscus plum ...