鹿茸基因组DNA提取方法的比较研究

Genomic DNA of Pilose Antler: Comparison of Extraction Methods

本研究旨在建立一种稳定、高效的鹿茸药材DNA提取方法,以满足后续的鹿茸DNA条形码鉴定方法需求。采用2种试剂盒、改良的CTAB法以及SDS法对市场上5个厂家共8批次的鹿茸药材进行基因组DNA的提取,参考2015版《中华人民共和国药典》四部通则9107考察了样品量、水浴温度、水浴时间3个因素。实验结果表明4种方法均可以从鹿茸药材中提取出基因组DNA,其中天根试剂盒提取效果最佳,改良的CTAB法提取效果较差;通过方法学考察得出使用天根试剂盒,提取条件为样品量70 mg、水浴温度56℃,水浴时间6 h时提取效果最佳。本研究优化得到的天根试剂盒提取鹿茸DNA方法操作简便,提取DNA浓度、质量优良,能够用于后续的DNA条形码鉴定中。

The objectives are to establish a stable and efficient method for extracting DNA from pilose antler and meet the needs of DNA barcode identification methods for pilose antler in the future. The genomic DNAs from 8 batches of velvet medicinal materials from 5 manufacturers on the market were extracted by two kinds of kits, modified CTAB method and SDS method. The three factors of sample size, water bath temperature and action time were investigated by referring to 9107 from the General Principles of Pharmacopoeia of the People’s Republic of China(Ⅳ, 2015). The results showed that all methods could extract genomic DNAs from pilose antler. TIANamp Genomic DNA Kit got the best extraction, while the modified CTAB method had worse extraction. By methodological study, TIANamp Genomic DNA Kit had the best extraction with the extraction condition of the sample quantity for 70 mg, and the water bath temperature 56℃ for 6 h. TIANamp Genomic DNA Kit screened in this study can use simple and convenient method to extract and obtain excellent concentration and quality DNA, so it can be used for subsequent identification of DNA barcode in the future.