黄鳝体表粘液中基因组DNA的提取与分析

黄鳝基因组DNA一直采用抽血或剪取肌肉来提取,但是采血或剪取肌肉后黄鳝成活率很低。为了保留亲本,尝试从黄鳝体表粘液中提取DNA,并采用紫外分光光度计对提取到的DNA的纯度和浓度进行了检测。结果表明,用蛋白酶K消化,酚仿抽提后从粘液中得到的DNA的纯度与从肌肉中提取到的相差不大,但是提取到的量较少。为了验证提取的DNA是不是黄鳝基因组DNA,用黄鳝actin引物和18S r RNA基因引物扩增之后检测到了粘液中的较亮的DNA片段。为了进一步确认检测到的是黄鳝的DNA,我们又对得到的DNA进行了扩增,并得到的产物进行了克隆和测序。结果表明,粘液中提取到的DNA是黄鳝的基因组DNA。随后又选择了60尾黄鳝,提取其粘液之后对其成活率进行统计,养殖1个月后黄鳝的成活率都在95%以上。此黄鳝体表粘液提取DNA方法能提取高质量的DNA,可用于用于PCR鉴定。

Extraction and analysis of genomic DNA from body surface mucus of ricefield eel (Monopterusalbus)

Genomic DNA of ricefield eel can be extracted from blood or muscle, but most of eels were dead afterwards. In order to preserve the sampling eels, we attempted to extract DNA from the body surface mucus, and the purity and concentration of the extracted DNA were detected by UV spectrophotometer. The results showed that no obvious difference between the DNA extracted from mucus and the DNA from the muscle. About 70% DNA was recovered after proteinase K and phenol treatment. In order to verify that the extracted genomic DNA is the ricefield eel DNA, we designed primers based on the actin and 18S rRNA genes and amplified one band from the mucus DNA. After being cloned and sequenced, the obtained product was the fragment of the actin gene. The results showed that the DNA extracted from mucus is the genomic DNA of ricefield eel. We then extracted mucus DNA from 60 eels and the survival rate of these eels was all above 95% after cultivating for one month.