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Broccoli SSR (simple sequence repeat) primer based on transcriptome sequencing and development

A transcriptome sequencing and cauliflower technology, applied in the biological field, can solve problems such as high cost, long time, and small number of SSR primers

Active Publication Date: 2018-01-19
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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Problems solved by technology

The development of traditional SSR markers is to design SSR primers based on the conserved regions on both sides of the gene sequence through the genome or downloading the EST sequence, which is a lot of work, long time, and expensive (Zhang et al., 2012), and the cauliflower in the NCBI database The number of EST sequences is limited and the number of SSR primers that can be developed is small

Method used

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  • Broccoli SSR (simple sequence repeat) primer based on transcriptome sequencing and development
  • Broccoli SSR (simple sequence repeat) primer based on transcriptome sequencing and development
  • Broccoli SSR (simple sequence repeat) primer based on transcriptome sequencing and development

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Embodiment Construction

[0015] 1 Materials and methods

[0016] 1.1 Transcriptome data sources

[0017] The cauliflower transcriptome data comes from the high-throughput transcriptome deep sequencing of cauliflower in this project. During sequencing, the cauliflower pedicels were taken and stored in liquid nitrogen for quick freezing, and then delivered to Beijing Biomicr for RNA-Seq transcriptome sequencing. The total data volume was 15.58 Gb, including 66,450 unigenes as background data for analysis.

[0018] 1.2 Plant material and its DNA extraction

[0019] The materials used for screening and verification of the designed primers were 24 cauliflower varieties collected and preserved by our research group (Table 1). The cauliflower DNA was extracted by the modified CTAB method (Lin Hui et al., 2015). Total RNA of each material was extracted with Trizol reagent (Invitrogen, USA) (Guo et al., 2009).

[0020] Table 1 Analysis materials of cauliflower SSR polymorphism

[0021]

[0022] 1.3 Tra...

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Abstract

The invention provides a broccoli SSR (simple sequence repeat) primer based on transcriptome sequencing and development. The broccoli SSR primer comprises 17 pairs of primers, wherein the sequences ofthe primers are shown in SEQ ID NO.1 to 34. The broccoli SSR primer has the advantages that a large amount of transcript data information is obtained by the transcriptome sequencing of the broccoli,a large amount of SSR primers is developed, the developed SSR marker is effectively demonstrated by the representative germplasm resources collected by a topic team, and the foundation is laid for thesubsequent study of genetic diversity, comparison and study of genomes, constructing of genetic spectrums, QTL (quantitative trait loci) location of important characters, utilization and storage of core resources, auxiliary breeding of molecules, and the like of the broccoli.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to cauliflower SSR primers developed based on transcriptome sequencing. Background technique [0002] brocoli( Brassica oleracea L. var. botrytis L. , 2n=2x=18) is a brassica vegetable in the genus Brassica of the family Brassicaceae, which is native to the Mediterranean coast (Liu Yunxia, ​​2011). Cauliflower is planted in Fujian, Zhejiang, Hubei and other regions of our country, and is one of the very important vegetables in our country. Cauliflower is rich in nutrition and has anti-cancer and anti-cancer effects. As a health-care vegetable with both medicine and food, the demand is increasing and the planting area is expanding (Zhu Shiyang et al., 2012). According to reports, my country's cauliflower ranks first in the world in terms of cultivation area and yield (Li Wenping et al., 2014). [0003] Research on cauliflower breeding in my country started relatively la...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 林珲朱海生李永平陈敏氡温庆放薛珠政叶新如
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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