基于简单加权分析的花椒果皮DNA快速提取方法比较研究

Comparative study on the rapid extraction methods of Zanthoxylum pericarp DNA based on simple additive weighting analysis

中文关键词:  花椒果皮  DNA提取  简单加权  快速提取

英文关键词:Zanthoxylum pericarp  DNA extraction  simple additive weighting  fast extraction

中文摘要:

      目的 开发和探究适用于花椒基因组DNA的快速提取方法。方法 选取8种不同品种的花椒, 分别利用3种不同的DNA提取方法, 包括一管式植物DNA抽提试剂盒、基于载体的纤维素滤纸提取法和无菌拭子浸入法, 进行花椒果皮基因组DNA快速提取, 比较不同DNA提取方法在花椒果皮基因组提取中的效果。其中纤维素滤纸提取法和无菌拭子浸入法均通过载体转移进行DNA的快速提取, 基于十六烷基三甲基溴化铵(hexadecyl trimethyl ammonium bromide, CTAB)和十二烷基硫酸钠(sodium dodecyl sulfate, SDS)两种不同的裂解液, 在3 min以内即可提取DNA并得到聚合酶链式反应(polymerase chain reaction, PCR)扩增体系。通过简单加权(simple additive weighting, SAW)分析, 基于PCR扩增能力、提取时间、提取成本、试剂安全性、程序简单性等方面进行综合评分。结果 所选的快速提取试剂盒不适用于花椒果皮这类含次生代谢物较多的实验材料, 而无菌拭子、纤维素滤纸两种载体提取得到的花椒果皮DNA均可以进行后续的PCR操作, 最终CTAB纤维素滤纸法以其低成本、短时间、高扩增效率等优势排名第一。结论 本研究探究了适用于花椒果皮基因组DNA的快速提取方法, 并且通过SAW分析得出CTAB纤维素滤纸法是最优选择, 表明载体转移法可以作为花椒基因组DNA快速提取方法之一, 为花椒基因组DNA快速提取提供了参考方案, 并为类似动植物基因组DNA的快速提取研究提供了参考。

英文摘要:

      Objective To develop and explore a rapid extraction method for genomic DNA of Zanthoxylum. Methods Eight different varieties of Zanthoxylum were selected, and 3 kinds of different DNA extraction methods, including one-tube plant DNA extraction kit, carrier-based cellulose filter paper extraction method and sterile swab immersion method, were used to quickly extract genomic DNA from the peel of Zanthoxylum, and the effects of different DNA extraction methods on the extraction of genomic DNA from the peel of Zanthoxylum were compared. Among them, the cellulose filter paper extraction method and the sterile swab immersion method were used to quickly extract DNA by vector transfer. Based on two different lysis buffers of hexadecyl trimethyl ammonium bromide (CTAB) and sodium dodecyl sulfate (SDS), DNA could be extracted in less than 3 minutes and a polymerase chain reaction (PCR) amplification system was obtained. Through simple additive weighting (SAW) analysis, comprehensive scoring was performed based on PCR amplification ability, extraction time, extraction cost, reagent safety, and program simplicity. Results The selected rapid extraction kit was not suitable for experimental materials containing more secondary metabolites such as Zanthoxylum pericarp, while the DNA extracted from the pericarp of Zanthoxylum by sterile swab and cellulose filter paper could be used for subsequent PCR operations, and finally, the CTAB cellulose filter paper method ranked first with its advantages of low cost, short time and high amplification efficiency. Conclusion In this study, a rapid extraction method suitable for the genomic DNA of Zanthoxylum pericarp is explored, and the CTAB cellulose filter paper method is the best choice through SAW analysis, indicating that the vector transfer method can be used as one of the rapid extraction methods for the genomic DNA of Zanthoxylum, which provides areference scheme for the rapid extraction of Zanthoxylum genomic DNA, and provides a reference for the rapid extraction of similar animal and plant genomic DNA.