CAJ | 학술논문

目的：建立简便、快捷、经济的模式小鼠总DNA提取方法，以快速鉴定大批量模式小鼠基因型。方法采用苯酚抽提法、异丙醇沉淀法、鼠耳煮沸法提取同种模式小鼠总DNA，对比DNA纯度、得率、耗费时间，并比较基因型鉴定结果。结果苯酚抽提法得率最高，异丙醇沉淀法最低；而纯度则按照苯酚抽提法、异丙醇沉淀法、鼠耳煮沸法顺序递减；在耗时上鼠耳煮沸法最短。三种方法提取的DNA均可做模版用于基因型鉴定。结论鼠耳煮沸法操作简单、成本最低，快速、基因型鉴定结果可靠，可用于规模化的基因型鉴定实验中。
목적：건립간편、쾌첩、경제적모식소서총DNA제취방법，이쾌속감정대비량모식소서기인형。방법채용분분추제법、이병순침정법、서이자비법제취동충모식소서총DNA，대비DNA순도、득솔、모비시간，병비교기인형감정결과。결과분분추제법득솔최고，이병순침정법최저；이순도칙안조분분추제법、이병순침정법、서이자비법순서체감；재모시상서이자비법최단。삼충방법제취적DNA균가주모판용우기인형감정。결론서이자비법조작간단、성본최저，쾌속、기인형감정결과가고，가용우규모화적기인형감정실험중。
Objective To establish a simple , fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice .Methods Three methods, i.e.phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C 57-rasmodel mice.The purity and yield of DNA obtained by the three methods were compared , and polymerase chain reaction ( PCR) assay was used to compare the efficacy of the three extraction methods .Results Among the three methods , phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield .In terms of DNA purity , the phenol extraction was the best and the mouse ear boiling method was the poorest .All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification .The time consumption of three methods are 12.5 hr ,13 hr and 0.18 hr.Mouse ear boiling method was significantly lower than that of the other two methods ( P <0.01 ) ,.The obtained total DNA can be stored at conventional -20℃for 7 days and 30 days later still can be used as a template for PCR reaction .Conclusions Among the three methods studied , the mouse ear boiling method is simple and with the lowest cost , so it is feasible for total DNA extraction in scaled genotyping experiments .