CAJ | 학술논문

以95%酒精保存的黄鳝(Monopterus albus)和斑鳢(Channa maculates)标本为材料,采用先沉降DNA再去除杂质的方法从鱼类标本中提取基因组DNA.基因组DNA的琼脂糖凝胶电泳和紫外分光光度法检测以及PCR扩增结果显示,本方法提取的鱼类基因组DNA的电泳主带清晰明亮;A_(260)/A_(280)值在1.7830-2.0144之间;PCR扩增产物条带清晰明亮,且单一整齐没有拖带,表明本方法可从酒精保存的鱼类标本中提取比较纯净的DNA,能够满足一般分子生物学试验需要.与传统苯酚/氯仿法相比,本方法操作简单快速,避免了苯酚等物质对后续实验的影响,可作为一种常规动物组织DNA提取方法.
이95%주정보존적황선(Monopterus albus)화반례(Channa maculates)표본위재료,채용선침강DNA재거제잡질적방법종어류표본중제취기인조DNA.기인조DNA적경지당응효전영화자외분광광도법검측이급PCR확증결과현시,본방법제취적어류기인조DNA적전영주대청석명량;A_(260)/A_(280)치재1.7830-2.0144지간;PCR확증산물조대청석명량,차단일정제몰유타대,표명본방법가종주정보존적어류표본중제취비교순정적DNA,능구만족일반분자생물학시험수요.여전통분분/록방법상비,본방법조작간단쾌속,피면료분분등물질대후속실험적영향,가작위일충상규동물조직DNA제취방법.
The genomic DNA of Monopaterus albus and Channa maculata preserved in 95%alcohol was extracted with a new method.Unlike traditional DNA isolation method,the present protocol precipitate DNA first,then remove impurities.The genomic DNA was appraised by agrose gel electrophoresis,spectrophotometer and PCR amplification.The results showed that the gel bands of genomic DNA were clear and bright without drawbands,and the ratio of A_(260)/A_(280) was between 1.7830 to 2.0144.It proved that this protocol could provide relatively pure DNAs which meet the needs of general molecular experiments.Furthermore,compared with the traditional method of phenol-chloroform,this protocol was not only fast and efficient,but also could avoid negative effects of residual phenol in the follow-up experiments.So this method might be used as a conventional protocol of extracting genomic DNAs from animal tissues.