摘要
分别采取PEG-6000沉淀法、酶直接消化法和超速离心法浓缩、纯化减蛋综合征病毒,并提取基因组DNA。结果表明:上述三种方法对1.5mL鸭胚尿囊液(HA≥14(log_2))提取的DNA量分别为148.4,189.2和210.4μg;其OD(260)/OD_(280)值分别为1.702,1.635,1.738,均在1.6～1.8之间,完全符合要求的纯度;经电泳显示,三种方法提取的DNA图谱也完全一致。提示,在无超速离心机的条件下,酶直接消化法和PEG-6000沉淀法提取的EDSV基因组DNA在纯度和数量方面,完全可以满足分子生物学的研究需要,且利用PEG-6000沉淀法,在时间、设备和试剂方面更为节简,可作为一般实验室操作时的首选方法。
PEG-6000 precipitation, direct proteolysis and ultra centrifugation were used to purify and extract genomic DNA(gDNA) from egg drop syndrome virus (EDSV) respectively. The results shows that the content of gDNA which is extracted from 1.5 mL EDSV allantoic liquid (HA≥14 (log2))is 148.4μg. 189.2μg and 210.4μg separately. The ratio of OD260 to OD280 is between 1.6 to 1.8, the electro photographs of gDNA are also the same. The quantity and purity of gDNA extracted by using PEG-6000 precipitation and direct proteolysis could meet the need with molecular research under the condition of no ultra centrifuge.
基金
河南省自然科学基金资助项目(0124040027)
