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| A New Method to Identify the Copy Number of Plasmid by Multiplex PCR |
| LI Yuan1,2, REN Chang-hong1, WU Yong-hong1, YE Qiao1, SHI Jin-ping1, QU Wu-bin1, ZHENG Xiao-fei1, LIU Hu-qi2, ZHANG Cheng-gang1,2 |
1. Beijing Institute of Radiation Medicine, State Key Laboratory of Proteomics, Beijing 100850, China;
2. The College of Life Science, Northwest Agriculture and Forest University, Yangling 712100, China |
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Abstract The PCR technique has been widely used in various fields of molecular biological studies. However, multiplex PCR for determination of plasmid copy number has not been reported. To further explore the research in this field, based on the multiple PCR primer design and evaluation computer programs, Two-plex PCR primers were disigned to amplify the bacterial genome and plasmid DNA; then using bacterial liquid of E. coli with different plasmid vectors as the template for multiple PCR reactions. Finally, the integrated optical density of the gel image was analyzed to determine relative copy number of the plasmids. The data showed that the result of relative copy number of the plasmids determined by multiplex PCR was consistent with which determined by Real-time PCR and Southern blot, indicating that multiplex PCR was an alternative and valuable method compared with traditional techniques in routine experiments. So a rapid and efficient method in identification of the plasmid copy number was established, which provides a suitable way to host cell selection by easily and rapidly determining the relative copy number of clone vector and expression vector.
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Received: 05 January 2011
Published: 28 June 2011
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Cite this article:
LI Yuan, REN Chang-hong, WU Yong-hong, YE Qiao, SHI Jin-ping, QU Wu-bin, ZHENG Xiao-fei, LIU Hu-qi, ZHANG Cheng-gang. A New Method to Identify the Copy Number of Plasmid by Multiplex PCR. China Biotechnology, 2011, 31(06): 93-98.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I06/93
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