唾液基因组dna提取和pcr扩增 - 第三军医大学学报.docVIP

一种从唾液中快速提取基因组DNA的方法和，林静1，陈龙辉2，张敏3，陈蔚文2（510006 广州，广东药学院基础学院1；510405 广州，广州中医药大学脾胃研究所2；510240 广州，广州市海珠区妇幼保健院中医科3） [摘要目的 探索一种快速、有效且对人体无的基因组DNA方法。方法碘化钾法和口腔拭子DNA提取试剂盒新鲜和室温放置一周的唾液标本的基因组DNA提取TP53、PRB-3基因PCR扩增。结果TP53和PRB-3基因结论School of Basic Courses, Guangdong Pharmaceutical University, 510006 Guangzhou; 2Pi-Wei Institute, Guangzhou University of Chinese Medicine, 510405 Guangzhou; 3Pediatrics of Traditional Chinese Medicine, Haizhu Maternal and Child health Hospital, 510240 Guangzhou) [Abstract] Objective To explore a fast, effective and noninvasive method of genomic DNA extraction. Methods Compare the genomic DNAs extracted by two DNA extraction methods of KI and kit from fresh and a week of salivary samples at room temperature, and PCR amplification for TP53 and PRB-3 genes from their extracted DNA. Ninety-four healthy children and adults were recruited and their salivary samplers were collected to check the stability of DNA extraction method based on KI. Resulsts Higher quailty genomic DNAs were extracted by the two methods of DNA extraction from fresh salivary samples although extracted DNA from a week of salivary samples at room temperature degradated obviously, and expectant DNA fragments were successfully amplified by PCR for TP53 and PRB-3 genes based on all these extracted DNAs. Although the genomic DNAs extracted by the mothod of KI from salivary samples of ninety-four healthy volunteers showed individual difference, DNA quality was stable and reliable. Conclusion Using the method of KI to extract genomic DNA from saliva was not only cheap and high performances, but also noninvasive for subjects, which was very suitability for large-scale epidemiological studies. [Key words] saliva; KI; DNA extraction; PCR amplification Supported by the National Natural Science Foundation of China (No., Science and Technology Planning Project of Guangdong Province (No. 2013A032500005) and Administration of Traditional Chinese Medicine of Guangdong Province in China (No.