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海洋沉积物DNA提取前的简易脱腐方法研究
引用本文:席峰,傅莲英,王桂忠,郑天凌.海洋沉积物DNA提取前的简易脱腐方法研究[J].高技术通讯,2006,16(5):539-544.
作者姓名:席峰  傅莲英  王桂忠  郑天凌
作者单位:1. 近海海洋环境科学国家重点实验室,厦门大学海洋与环境学院,福建,361005;集美大学水产生物技术研究所,福建,361021
2. 厦门大学生命科学学院,福建,361005
3. 近海海洋环境科学国家重点实验室,厦门大学海洋与环境学院,福建,361005
4. 近海海洋环境科学国家重点实验室,厦门大学海洋与环境学院,福建,361005;厦门大学生命科学学院,福建,361005
基金项目:国家科技攻关项目 , 中国科学院资助项目 , 教育部长江学者和创新团队发展计划 , 福建省科技厅科研项目
摘    要:针对海洋沉积环境样品DNA提取中的腐殖酸去除难题,采取先脱除腐殖酸再提取DNA的策略,进行了海洋沉积物DNA提取前的简易脱腐方法研究.依据腐殖酸的理化性质, 遴选出由Tris-HCl、EDTA、Na4P2O7、NaCl、PVP、Triton X-100及脱脂奶粉组成脱腐缓冲液,有效地脱除了腐殖酸.之后采用温和的溶菌酶-蛋白酶K-SDS直接裂解法,获取了大片段(分子量21kb以上)可进行rpoB 基因PCR扩增的海洋沉积物DNA,为海洋沉积环境分子生态学研究与海洋生物活性物质开发奠定了基础.

关 键 词:海洋  沉积物  DNA  腐殖酸  脱除
收稿时间:2005-09-08
修稿时间:2005年9月8日

A simple method for removing humic acids from marine sediment samples prior to DNA extraction
Xi Feng,Fu Lianying,Wang Guizhong,Zheng Tianling.A simple method for removing humic acids from marine sediment samples prior to DNA extraction[J].High Technology Letters,2006,16(5):539-544.
Authors:Xi Feng  Fu Lianying  Wang Guizhong  Zheng Tianling
Affiliation:State Key Laboratory of Marine Environmental Science, College of Oceanography and Environmental Science, Xiamen University, Xiamen 361005; School of Life Sciences, Xiamen University, Xiamen 361005; Aquaculture Biological Technique Institute, Jimei University, Xiamen 361021
Abstract:A strategy of removing humic acids (HAs) prior to DNA extraction was conducted to resolve the problem of removing HAs from marine sediments in DNA extraction. According to the physical and chemical properties of HAs, the HAs' removing buffer composing of Tris-HCl, EDTA, Na_4P_2O_7, NaCl, PVP, Triton X-100 and skim milk was screened out, which washed HAs out from the marine sediment samples successfully. Then a mild lysozyme-proteinase K-SDS direct lysis method was applied and the large fragment DNA was recovered from the marine shrimp pond, seaport and mangrove sediments samples, the molecular mass of which is more than 21 kb. These DNA extractions contain the sediment DNA which could be PCR amplified for rpoB gene. These work laid a foundation for molecular ecology research and exploiting the bioactive substances from marine sediments.
Keywords:marine  sediment  DNA  humic acids (HAs)  remove
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