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PCR reaction system for SCoT molecular marker of lucuma nervosa

A technology of reaction system and molecular markers, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., to achieve the effect of strong applicability, high applicability, great scientific value and application value

Active Publication Date: 2022-04-08
SOUTH ASIAN TROPICAL AGRI SCI RES INST OF GUANGXI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no literature reports on the use of SCoT molecular markers for quality resource identification and genetic diversity analysis of lucuma at home and abroad. Therefore, constructing and optimizing the SCoT-PCR reaction system of lucuma and obtaining the best reaction system will provide a basis for the use of SCoT molecular markers in the future. Carry out research on genetic diversity analysis, germplasm protection and innovative selection of lucuma germplasm resources to provide a technical support basis

Method used

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  • PCR reaction system for SCoT molecular marker of lucuma nervosa
  • PCR reaction system for SCoT molecular marker of lucuma nervosa
  • PCR reaction system for SCoT molecular marker of lucuma nervosa

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Embodiment

[0023] 1. Experimental materials:

[0024] The samples of the present invention are all collected from Guangxi South Subtropical Agricultural Science Research Institute, wherein No. 1 is "Xiantao No. 1" bred by Guangxi South Subtropical Agricultural Science Research Institute, and the remaining samples are lucuma from different regions in China.

[0025] 2. Experimental content:

[0026] 1. Extraction and detection of lucuma DNA

[0027] The young leaves of lucuma were taken, DNA was extracted from the sample with a kit, and the DNA concentration, OD260 / 280 and OD260 / 230 were detected by NanoDrop; the integrity of the sample DNA was detected by agar gel electrophoresis.

[0028] 2. Optimization of lucuma SCoT-PCR reaction system

[0029] The optimization experiment was carried out with sample No. 1 and primer SCoT1 (5'-CAACAATGGCTACCACCA-3'). DNA, Mg 2+ , dNTPs, primers and Taq polymerase 5 influencing factors are set to 4 different levels (see Table 1 for different factor...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) system for a SCoT molecular marker of lucuma nervosa, and belongs to the technical field of biology. The PCR reaction system comprises the following components: 80ng of template DNA (deoxyribonucleic acid), 3mmol / L of Mg < 2 + >, 0.25 mmol / L of dNTPs (deoxyribonucleoside triphosphates), 0.6 mu mol / L of primer and 2U of Taq enzyme. The primer is any one of SCoT9, SCoT16, SCoT24, SCoT33 and SCoT48, and the nucleotide sequence of the primer is as shown in SEQ ID NO. 1 to SEQ ID NO. 5. The strip amplified by the established SCoT-PCR reaction system of the lucuma nervosa is high in stability and high in definition, has the advantages of simplicity in operation, high sensitivity, good repeatability and the like, and can realize the analysis of genetic diversity of the lucuma nervosa.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the construction of an optimal PCR reaction system for lucuma SCoT molecular markers. Background technique [0002] Lucuma nervosa (Lucuma nervosa A.DC), also known as lion head fruit, egg fruit, peach olive, fairy peach, etc., is a perennial tropical evergreen rare fruit tree of the genus Lucuma in the family Sapotaceae. Native to Cuba and tropical America, it was introduced to China in the 1930s and is suitable for planting in areas south of 24° north latitude in my country (Deng Chengju et al., 2013). The flesh of lucuma is yellow and powdery, like cooked egg yolk. It has a unique taste and is rich in essential amino acids. From December to March of the next year, it is an off-season fruit with a long fruit supply period, which has a good regulating and promoting effect on the fruit market (preliminary report on the introduction and planting of lucuma, Luo Xinping, 20...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCY02A50/30
Inventor 周彩霞李文砚周婧卢美瑛韦雪英韦优赵静蒋娟娟黄丽君卓福昌唐景美周之珞颜桢灵罗培四杨志强卢艳春孔方南
Owner SOUTH ASIAN TROPICAL AGRI SCI RES INST OF GUANGXI
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