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发明公开
CN102994493A 一种海参体壁总DNA提取方法 无效 - 驳回

一种海参体壁总DNA提取方法
摘要：
本发明公开了一种海参体壁总DNA提取方法。它包括样品预处理、样品的裂解、样品消化液用氯仿：异戊醇抽提，含有DNA的水相加入等体积的异丙醇高盐溶液，-20℃下放置20min，12000-13000g/min离心12-15min，弃上清，取DNA沉淀；所述的异丙醇高盐溶液是由异丙醇和5mol/L NaCl按体积比3：1混合而成；用体积分数70%乙醇水溶液的洗涤DNA沉淀，然后12000-13000g/min离心3-5min，弃上清；重复操作2-3次，将DNA沉淀干燥，再用0.1×TE溶液充分溶解保存。因此利用本发明的海参体壁总DNA提取方法能够高质量的提取到海参DNA，其DNA纯度高，避免了粘多糖、胶原蛋白和色素等影响，能满足后续PCR等分子生物学实验的要求。
摘要（英）：

The invention discloses a method for extracting total DNA (deoxyribonucleic acid) from body wall of sea cucumber. The method comprises the steps: pretreating samples, cracking samples, extracting sample digestive fluid with chloroform and isoamyl alcohol, adding isometric isopropyl alcohol high-salt solution into DNA-containing water phase, standing for 20min at minus 20 DEG C, centrifuging for 12-15min at 12000-13000g/min, discarding supernatant, and taking DNA precipitate, wherein the isopropyl alcohol high-salt solution is prepared by mixing isopropyl alcohol and 5mol/L NaCl according to the volume ratio of 3:1; washing the DNA precipitate with ethanol water solution the volume fraction of which is 70%, then centrifuging for 3-5min at 12000-13000g/min, and discarding supernatant; and repeating the operations by twice to three times, drying the DNA precipitate, then fully dissolving with 0.1*TE solution, and storing. Therefore, the DNA of the sea cucumber can be extracted in a high-quality manner by adopting the method for extracting total DNA from body wall of sea cucumber, the extracted DNA is high in purity, the influence from mucopolysaccharide, collagen, pigment and the like can be avoided, and the requirements on the subsequent molecular biology experiments such as PCR (Polymerase Chain Reaction) can be met.

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