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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. As a result of the large amount of fats, proteins and polysaccharides, Hashimoto’s matrix severely interferes with the extraction of genomic DNA. Genomic DNA was extracted from Cortex clavatus by CTAB method, Enzymatic detergent method, Sodium dodecylbenzenesulfonate (SDS) method and SDS-CTAB method. The UV-Vis spectrophotometry , Gel electrophoresis and Co Ⅰ sequence amplification method to compare the DNA quality obtained by four extraction methods. The results showed that DNA extracted by SDS-PAGE, CTAB and SDS-PAGE combined with CTAB method was the best. By comparing with DNA barcodes of the original species, it was proved that all the three methods could provide the required DNA, which laid the foundation for the molecular biology research of the snow clams.