Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (24): 4430-4434.doi: 10.3969/j.issn.1673-8225.2010.24.014

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Comparison of four methods for DNA extraction from paraffin-embedded tissues

Yuan Ya-ting1, Jiang Yue-xin1, Yin Xiao-wen2, Jiang Xing-tang2   

  1. 1 Medical College of Xiamen University, Xiamen  361004, Fujian Province, China; 2 Department of Respiratory Medicine, Zhongshan Hospital, Xiamen University, Xiamen  361004, Fujian Province, China
  • Online:2010-06-11 Published:2010-06-11
  • Contact: Jiang Xing-tang, Professor, Department of Respiratory Medicine, Zhongshan Hospital, Xiamen University, Xiamen 361004, Fujian Province, China xmzsjxt@163.com
  • About author:Yuan Ya-ting★, Studying for master’s degree, Medical College of Xiamen University, Xiamen 361004, Fujian Province, China yyting_123@163.com

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Abstract:

BACKGROUND: Because of the damage of formalin-fixed paraffin-embedded tissue (FFPET) to DNA during the production and preservation, it is difficult to extract high quality and sufficient DNA for the following researches such as polymerase chain reaction (PCR). Therefore, it is essential and urgent to investigate an effective and economic method for extraction of DNA from FFPET.
OBJECTIVE: To investigate the influence of four DNA extraction methods for FFPET on DNA quality, and to identify the optimal method for DNA extraction from FFPET.
METHODS: Twenty formalin-fixed, paraffin-embedded non-small cell lung cancer (NSCLC) tissue specimens were selected and divided into four groups according to the method for DNA extraction. Four methods included xylene dewaxing and phenol-chloroform method, improved TES dewaxing with water-bath and phenol-chloroform method, genomic DNA extraction kit, improved TES dewaxing with water-bath and genomic DNA extraction kit. The quality of obtained DNA was analyzed by electrophoresis and PCR amplification.
RESULTS AND CONCLUSION: For paraffin-embedded tissue specimens, better DNA fragments could be obtained by using the improved TES dewaxing with water-bath and phenol-chloroform method and improved TES dewaxing with water-bath and genomic DNA extraction kit. The A260/A280 ratio of DNA obtained by using improved TES dewaxing with water-bath and phenol-chloroform method and improved TES dewaxing with water-bath and genomic DNA extraction kit were statistically different from genomic DNA extraction kit (P < 0.01). There was no significant difference in efficiency among four mehtods (P > 0.1). The aim trips were as bright as the positive control when using the DNA extraction by improved TES dewaxing with water-bath and phenol-chloroform method as the template. The results demonstrated that improved TES dewaxing with water-bath and phenol-chloroform method is an optimal method for extraction of DNA from FFPET.

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