Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (15): 2820-2822.doi: 10.3969/j.issn.1673-8225.2012.15.035

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Comparison of three methods for preparing DNA sequencing templates 

Qin Chuan1, Zhang Jun-he2, Robert A. Mcknight3○, Liu Shi-guo1   

  1. 1Laboratory of Molecular Biology, 2Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang  453003, Henan Province, China; 3Medicine University of Utah, Salt Lake City  84101, USA
  • Received:2011-09-25 Revised:2011-10-20 Online:2012-04-08 Published:2012-04-08
  • Contact: Liu Shi-guo, Doctor, Professor, Laboratory of Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China lsg@xxmu.edu.cn
  • About author:Qin Chuan★, Master, Lecturer, Laboratory of Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China huaxingene@163.com

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Abstract:

BACKGROUND: The quality of DNA templates plays a crucial role in DNA sequencing.
OBJECTIVE: To provide an economical, simple method for genomic DNA sequencing and methylated DNA sequencing.
METHODS: Plasmid extraction was performed using NucleoSpin multi-96 plus plasmid kit and edgebio kit SeqPrep™ 96 HP plasmid prep system, respectively. Primer pairs containing target fragments were designed for the plasmids. PCR products were purified after amplification. DNA sequencing templates were prepared by the three methods above.
RESULTS AND CONCLUSION: There was no significant difference in the genomic DNA sequencing results between the three methods used in the experiment (P > 0.05). As for the methylated DNA sequencing, the NucleosPin multi-96 plus plasmid kit was better than the other two methods (P < 0.05). These findings indicate that the three methods are suitable for genomic DNA sequencing, while the NucleosPin multi-96 plus plasmid kit is more situable for the methylated DNA sequencing.
 

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