湖北农业科学 ›› 2023, Vol. 62 ›› Issue (9): 170-174.doi: 10.14088/j.cnki.issn0439-8114.2023.09.030

• 生物工程 • 上一篇    下一篇

基于实时荧光定量PCR的番木瓜多重PCR高效检测方法

张玉山, 陈洁彬, 吴亮君, 朱璇, 欧阳淑芬, 沈志华   

  1. 电子科技大学中山学院材料与食品学院,广东 中山 528402
  • 收稿日期:2022-08-06 出版日期:2023-09-25 发布日期:2023-10-24
  • 作者简介:张玉山(1966-),男,黑龙江密山人,教授,博士,主要从事生物技术及现代农业绿色可持续发展研究,(电话)15113354163(电子信箱)class2007ok@163.com。
  • 基金资助:
    广东省科技发展专项资金“大专项+任务清单”项目(2019-319; 2020sdr009)

Efficient detection method of papaya multiple PCR based on real time fluorescence quantitative PCR

ZHANG Yu-shan, CHEN Jie-bin, WU Liang-jun, ZHU Xuan, OUYANG Shu-fen, SHEN Zhi-hua   

  1. College of Materials and Food, Zhongshan Institute, University of Electronic Science and Technology of China, Zhongshan 528402, Guangdong, China
  • Received:2022-08-06 Online:2023-09-25 Published:2023-10-24

摘要: pCaMV35S基因核酸检测试剂盒反应体系为基础,添加多重PCR反应所需的目标基因引物和番木瓜(Carica papaya L.)DNA,进行多重PCR反应,同时考察不合成荧光探针的情况下对目标基因进行实时荧光定量检测。结果表明,在实时荧光定量PCR反应体系中,多重PCR扩增的基因条带比常规PCR扩增的基因条带更亮,在不合成荧光探针的情况下实现目标基因的实时荧光定量PCR反应,以期建立一种基于实时荧光定量PCR的多重PCR检测方法。

关键词: 转基因番木瓜(Carica papaya L.), 多重PCR反应, 实时荧光定量PCR, 荧光探针, pCaMV35S基因核酸检测试剂盒(PCR-荧光探针法)

Abstract: Based on the pCaMV35S gene nucleic acid detection kit reaction system, the target gene primers and papaya(Carica papaya L.) DNA required for multiple PCR reactions were added to conduct multiple PCR reactions. At the same time, the effect of real-time fluorescence quantification of the target gene without synthesizing fluorescent probes was investigated. The results showed that in the real-time fluorescence quantitative PCR reaction system, the gene bands amplified by multiple PCR were brighter than those amplified by conventional PCR, achieving real-time fluorescence quantitative PCR reaction of target genes without synthesizing Fluorescence probes, in order to establish a real-time fluorescence quantitative PCR detection method based on real time fluorescence quantitative PCR.

Key words: transgenic papaya (Carica papaya L.), multiple PCR reactions, real time fluorescence quantitative PCR, fluorescence probe, PCaMV35S gene nucleic acid detection kit (PCR fluorescence probe method)

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