雪蛤基因组DNA提取方法的研究及条形码分子鉴定

DNA extraction from Ranae oviductus and molecule barcode identification

高质量的DNA是开展分子生物学研究的基础,雪蛤基质由于含有大量脂肪、蛋白质和多糖,严重干扰其基因组DNA的提取。采用十六烷基三甲基溴化铵(CTAB)法、加酶洗洁精法、十二烷基苯磺酸钠(SDS)法、SDS结合CTAB法提取雪蛤基因组DNA,通过紫外分光光度法、凝胶电泳法及CoⅠ序列扩增法比较了4种提取方法得到的DNA质量。结果显示,加酶洗洁精法、SDS法、SDS结合CTAB法均可提取到雪蛤DNA,其中以SDS结合CTAB法获得的DNA质量最佳。通过与基原物种的DNA条形码进行比对,证明3种方法均可提供符合要求的DNA,为雪蛤的分子生物学研究奠定了基础。

High quality DNA is the basis of molecular biology research. Due to the rich content of fats, proteins and polysaccharides, extraction of the genomic DNA is tough for Ranae oviductus. Four DNA extraction methods were established for Ranae oviductus, as cetyltrimethyl ammonium bromide（CTAB） method, detergent adding enzyme method, sodium dodecyl sulfate（SDS） method, SDS combined with CTAB method. Compared the genomic DNA quality extracted from four methods by ultraviolet spectrophotometric method, gel electrophoresis and Co Ⅰ sequence amplification. Results showed that detergent adding enzyme method, SDS method, SDS combined with CTAB method could produce genomic DNA, among them, DNA from SDS combined with CTAB method was best. By comparison with basic species, the established extraction methods were proved to be suitable for DNA barcoding research of Ranae oviductus. The research provided the basis for molecular biology research of Ranae oviductus.