6种食用香辛料基因组DNA提取方法比较

Comparison of Six Methods for Extraction of Genomic DNA from Edible Spices

采用TIANGEN植物基因组DNA提取试剂盒法、TIANGEN DNAsecure新型植物基因组DNA提取试剂盒法、QIAGEN DNeasy plant kit法、Nucleospin®Food kit法、改良十六烷基三甲基溴化铵(cetyltrimethylammonium bromide,CTAB)法和十二烷基硫酸钠(sodium dodecyl sulfate,SDS)-CTAB法6种方法对来自植物根、茎、叶、花蕾、果实及种子的19种常见食用香辛料的DNA进行提取,并比较各方法的提取效果。结果表明,QIAGEN DNeasy plant kit法和TIANGEN植物基因组DNA提取试剂盒法所得DNA的聚合酶链式反应(polymerase chain reaction,PCR)扩增成功率最高,但QIAGEN DNeasy plant kit法提取的DNA质量浓度低,不利于后续研究。本研究建议将TIANGEN植物基因组DNA提取试剂盒法作为食用香辛料基因组DNA提取的优选方法,并用该方法进行市售香辛料粉的DNA提取。结果显示,所有市售香辛料粉的DNA质量浓度、纯度及PCR扩增效率均符合实验要求。表明TIANGEN植物基因组DNA提取试剂盒法适合食用香辛料DNA提取,而对于皮类及坚硬的食用香辛料应增加样品量及裂解时间。

In this study,we compared six different DNA extraction methods,including TIANGEN plant genomic DNA kit,TIANGEN DNAsecure plant kit,QIAGEN DNeasy plant kit,Nucleospin®Food kit,modified cetyltrimethylammonium bromide(CTAB)method,and sodium dodecyl sulfate(SDS)-CTAB method by examining their effectiveness for extracting DNA from 19 edible spices from plant roots,stems,leaves,buds,fruits,and seeds.The results showed that the DNAs obtained by the QIAGEN DNeasy plant kit and the TIANGEN plant genomic DNA kit were most successfully amplified by polymerase chain reaction(PCR).However,the QIAGEN DNeasy plant kit yielded a low DNA concentration,being unfavorable for subsequent studies.Therefore,we recommended that the TIANGEN plant genomic DNA kit should be preferably used for the extraction of genomic DNA from edible spices and DNA from commercial spice powder.The results showed that the concentration,purity and PCR amplification efficiency of DNA extracted from all commercial spice powders met the experimental requirements.Moreover,the sample amount and lysis time should be increased for DNA extraction from bark and hard edible spices using the TIANGEN plant genomic DNA kit.