麦冬基因组DNA提取方法研究

Study on Methods of Genomic DNA Extraction from Lilyturf

为了从富含多糖、多酚的顽拗植物麦冬中分离出高质量基因组DNA,分别建立了改良CTAB法和改良SDS法。通过使用核分离液和CTAB/NaCl溶液、提取液中加入0.35倍体积无水乙醇等最大限度地除去杂质。将两种方法提取的8种麦冬DNA与两种离心柱式试剂盒提取的DNA在纯度、产率等方面进行对比,并进行双酶切和SRAP分析。麦冬类植物SRAP反应体系的建立尚属首次。结果表明,改良CTAB法和改良SDS法均能从8种麦冬叶片中提取到高质量的基因组DNA,改良CTAB法提取纯度更高,提取幼叶DNA纯度可以与离心柱式试剂盒媲美,能够满足多种分子生物学试验要求。在后续试验中,用改良CTAB法从20多种沿阶草族植物中提取到了高纯度DNA。该方法通用性好,提取的DNA纯度高,对其他顽拗类植物基因组DNA的提取具有借鉴意义。

In order to isolate high quality genomic DNA from lilyturf included in recalcitrant plants rich in polysaccharides and polyphenols,the improved CTAB method and the improved SDS method were respectively established in this study.In order to removing the impurity furthest,nuclear isolation buffer and CTAB/NaCl solution were used and 0.35 volume absolute ethanol was added in extracting solution in the extraction protocols.The DNA extracted from 8 Lilyturf by these two methods was compared with DNA extracted by two DNA extraction column kit in purity,yield,etc,double digestion and SRAP analysis.It was the first time that the SRAP reaction system of Lilyturf was established.The results showed that high quality genomic DNA was separately extracted from leaves of 8 lilyturf by the improved CTAB method and the improved SDS method.The DNA extracted by the improved CTAB method was purer.The purity of DNA extracted by the improved CTAB method was comparable to that of DNA extracted by these two kits,meeting the requirements of varieties of molecular biology experiments.In the following experiment,high quality genomic DNA had been extracted from twenty more species of Ophiopogoneae plants by the improved CTAB method.This method had good generality,high purity DNA,and provided satisfactory reference to extract genomic DNA from other recalcitrant plants.