槟榔叶表微生物基因组DNA提取方法的比较

Comparison of Microbial Genomic DNA Extraction Methods from Areca catechu Leaf Surface

为确定槟榔叶表微生物基因组DNA的最佳提取方法,本研究采用改良CTAB法、MP Biomedicals土壤试剂盒、QIAGEN土壤试剂盒、天根多糖多酚植物试剂盒和MP Biomedicals植物试剂盒分别提取槟榔叶表微生物基因组DNA,通过超微量分光光度计测定、变性梯度凝胶电泳和聚合酶链式反应(PCR)对提取效果进行比较。结果表明,采用MP Biomedicals植物试剂盒提取的DNA效果最好,DNA浓度为81.57μg/mL,A_(260)/A_(280)值为1.80,A_(260)/A_(230)值为2.06,PCR扩增产物亮度高、无杂带,且具有提取时间短、效率高、污染小等特点。本研究为槟榔叶表微生物基因组DNA的提取提供了参考,为后续进行分子生物学实验提供了科学依据。

In order to determine the best extraction method of microbial genomic DNA from Areca catechu leaf surface,the modified CTAB method,SPINeasy DNA Kit for Soil(MP Biomedicals),DNeasy PowerSoil Pro Kit(QIAGEN),Super Plant Genomic DNA Kit(Polysacchardes&Polyphenolics-rich)(TIANGEN)and SPINeasy DNA Kit for Plant(MP Biomedicals)were utilized to extract microbial genomic DNA from Areca catechu leaf surface.,The extraction results were compared by ultrafine spectrophotometer,denaturation gradient gel electrophoresis and polymerase chain reaction(PCR).The result showed that SPINeasy DNA Kit for Plant(MP Biomedicals)has the best effect on DNA extraction,with DNA concentration of 81.57μg/mL,A_(260)/A_(280)value of 1.80 and A_(260)/A_(230)value of 2.06.The PCR amplified product has high intensity and no stray bands.It has the characteristics of short extraction time,high efficiency and minimal contamination.This study provides a reference for the extraction of genomic DNA from Areca catechu leaf surface,and provides a scientific basis for the subsequent molecular biology experiments.