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由于南极鱼的体内存在抗冻蛋白,不同于一般鱼类,常规基因组DNA的提取较为困难。为建立南极鱼基因组DNA提取的有效方法,利用贝氏肩孔南极鱼(Trematomus bernacchii)为材料,采用改进的酚-氯仿法提取基因组DNA,与传统酚-氯仿法和快速试剂盒抽提法作对比,并通过琼脂糖凝胶电泳、紫外分光光度计测定、以及PCR扩增等手段进行基因组DNA质量的检测。结果表明:利用改进的基因组DNA提取方法,从贝氏肩孔南极鱼获得的基因组DNA,电泳条带整齐明亮,A260/A280值接近1.80,适合后续的PCR扩增实验,可满足基于PCR技术的相关分子生物学实验需要。
Due to the presence of antifreeze proteins in Antarctic fish, unlike conventional fish, conventional genomic DNA extraction is more difficult. In order to establish an effective method for extracting genomic DNA of Antarctic fish, genomic DNA was extracted by using the improved phenolic-chloroform method with Trematomus bernacchii as material. Compared with traditional phenol-chloroform method and rapid kit extraction The quality of genomic DNA was detected by agarose gel electrophoresis, UV spectrophotometer and PCR amplification. The results showed that genomic DNA obtained from the Antarctic fish of Bayesian gland with neat and bright electrophoresis bands and the value of A260 / A280 approaching 1.80, which is suitable for subsequent PCR amplification experiments, could meet the needs of PCR-based Related molecular biology experiments need.