Oncidium EST-SSR labeling primer and application thereof

A technology for labeling primers and oncidium, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.

Active Publication Date: 2016-01-06
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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  • Oncidium EST-SSR labeling primer and application thereof
  • Oncidium EST-SSR labeling primer and application thereof
  • Oncidium EST-SSR labeling primer and application thereof

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[0055] The 10 oncidium EST-SSR marker primer pairs of the present invention are obtained by the following method:

[0056]Oncidium EST sequences were downloaded from the EST database in NCBI, and the EST sequences containing SSR sites were searched manually using SSRIT online software. The search criteria are: the minimum number of repetitions of two, three, four, five and six nucleotide motifs is 5 repetitions, and the SSR site information of each site is searched in turn. Use Primer5.0 software to design PCR amplification primers. The main parameters of primer design are: primer length 18-22bp; annealing temperature 48°C-54°C; difference between upstream and downstream primer annealing temperatures not higher than 3°C; GC content 35% ~60%; the expected fragment of the PCR amplification product of Oncidium EST-SSR is between 150~500bp. Synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0057] The present invention provides 10 Oncidium molecular mar...

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Abstract

The invention provides an Oncidium EST-SSR labeling primer and application thereof. The primer comprises a primer pair as follows: 5'-ATCGTAATCCTGAAGCGTATC-3' and 5'-AAGCCCAAACTATTCCATT-3'. The application of the Oncidium EST-SSR labeling primer comprises the steps of carrying out PCR amplification by taking a DNA sample extracted from Oncidium plants as a template, and labeling Oncidium molecules, wherein a reaction system includes the following components by volume (20mu L in all): 11.3mu L of ddH2O, 2mu L of 10*Buffer, 2mu L of 2.5mmol.L<-1> dNTPs, 1 microliter of each of 10mu mol.L<-1> primers in the primer pair, 0.2mu L of 5U.mu L<-1> Taq polymerase and 2.5mu L of 20ng/mu L template DNA.

Description

technical field [0001] The invention specifically relates to an Oncidium EST-SSR labeled primer and application thereof. Background technique [0002] Oncidium (Oncidium), also known as dancing orchid, is an important pot-cut dual-purpose flower. As a commodity, oncidium occupies a certain share in the world's flower market. Oncidium has developed rapidly since its discovery, and it has been produced on a large scale in many countries such as Thailand, Singapore, and Malaysia. Oncidium in a broad sense is the general term for the genus Oncidium and its related genera. Modern cultivars are all hybrids between species of Oncidium and related genera within subfamilies. The genetic background is very complicated. Therefore, research Its genetic diversity is of great significance. Since the 1990s, my country has vigorously developed the oncidium industry in Guangdong, Hainan, Fujian and other places. At present, the research on Oncidium in my country mainly focuses on tissue c...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 黄敏玲林榕燕钟淮钦罗远华祁世明
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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