六种试剂盒提取血液基因组DNA效果比较  

Comparison of DNA Extraction Effects by 6 Blood Genomic DNA Extraction Kits

作　　者：张艳[1] 王金鸿[1] 吕亚莉[1] 菅复春[1] 张龙现[1] 宁长申[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002

出　　处：《中国草食动物科学》2014年第2期9-12,共4页China Herbivore Science

基　　金：国家现代肉羊产业技术体系建设专项(nycytx-39)

摘　　要：为筛选一种高效、快捷的血液基因组DNA提取试剂盒,采集24份新鲜西藏绵羊血液样本,选用6种市售商业化试剂盒(LifeFeng柱式试剂盒DK601和非柱式试剂盒DK602,康为世纪非柱式试剂盒CW0544和柱式试剂盒CW0546,上海生工非柱式试剂盒SK8224和柱式试剂盒SK8254),比较所提DNA的浓度、纯度、得率以及对嗜吞噬细胞无形体16S rRNA进行巢式PCR扩增,综合评价不同试剂盒DNA提取效果。结果表明,应用LifeFeng柱式试剂盒DK601提取的DNA浓度和纯度均最高,得率较高,分别达到75.63 ng/μL、1.894(OD260/OD280)、18.91 ng/μL;且嗜吞噬细胞无形体16S rRNA基因的PCR扩增条带清晰度最优。因此,LifeFeng DK601可作为提取绵羊血液基因组DNA及嗜吞噬细胞无形体(Anaplasma phagocytophilum)病原鉴定的首选试剂盒。In order to select a relatively better DNA extraction kit for the extraction of genomic DNA, fresh blood samples were taken from 24 Tibetan sheep, and 6 commercial kits,including the LifeFeng DNA extraction kit DK601 and 602, CWBIO DNA extraction kit CW0544 and CW0546, Sangon DNA extraction kit SK8224 and SK8254, were chosen as the DNA extraction reagents. The qualities of extracted DNAs were evaluated by determination of the concentrations, the purities and extraction rates, as well as the effects of nested PCR targeted for the Anaplasma phagocytophilum 16S rRNA gene. The results showed that the highest DNA concentration, purity and extraction rate, which came up to 75.63 ng/μL, 1.894 （OD260/OD280）and 18.91 ng/μL, respectively, could been achieved by employing the LifeFeng DNA extraction kit DK601. Furthermore, the best results of PCR re- action were observed using the DNA samples harvested from this kit. It therefore been concluded that the LifeFeng DNA extraction kit DK601 was the relatively better reagent for the genomic DNA extraction and detection ofA naplasmaphagocytophilum from the blood in sheep research.

关 键 词：DNA提取试剂盒 血液基因组DNA PCR 

分 类 号：S826.2[农业科学—畜牧学]