Designation and Preparation of Fluorescently Labeled DNA Internal Lane Standard
Author(s): Yang Pian, Wang Bangyi, Ye Jian, Liang Jingqing, Zhao Xingchun, School of Forensic Medicine, Shanxi Medical University, Institute of Forensic Science, Ministry of Public Security
Pages: 56-58
Year: 2014 Issue: 6
Journal: Life Science Instruments
Keyword: Fluorescently labeled; pMD18-T Vector; PCR; Internal lane standard;
Abstract: Objective: To prepare the fluorescently labeled DNA internal lane standard range from 65 bp to 500 bp. Method: Clone and double digestion methods were employed to prepare the template of p MD18-T vector ligated a recovery DNA fragment, a fixed and fluorescently labeled forward primer and common reverse primers were designed, and PCR products were analyzed by PCR amplification and ABI 3100 Genetic Analyzer. Result: The size of PCR products is 65bp、105bp、149bp、200bp、241bp、269bp、311bp、345bp、400bp、450bp、500bp,in line with the expected fragment size. Conclusion: The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.
Pages: 56-58
Year: 2014 Issue: 6
Journal: Life Science Instruments
Keyword: Fluorescently labeled; pMD18-T Vector; PCR; Internal lane standard;
Abstract: Objective: To prepare the fluorescently labeled DNA internal lane standard range from 65 bp to 500 bp. Method: Clone and double digestion methods were employed to prepare the template of p MD18-T vector ligated a recovery DNA fragment, a fixed and fluorescently labeled forward primer and common reverse primers were designed, and PCR products were analyzed by PCR amplification and ABI 3100 Genetic Analyzer. Result: The size of PCR products is 65bp、105bp、149bp、200bp、241bp、269bp、311bp、345bp、400bp、450bp、500bp,in line with the expected fragment size. Conclusion: The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.
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