金柑叶片和果实总DNA提取方法比较

摘要

为了研究不同提取方法对金柑总DNA提取质量的影响,以融安金柑等3个金柑品种的叶片和果实为材料,采用SDS提取法、CTAB提取法、试剂盒法提取金柑总DNA,并对所提取DNA的总浓度、A260/280、A260/230、凝胶电泳结果、SCoT扩增产物进行比较分析.结果表明:CTAB提取法和试剂盒提取法能较好地去除金柑叶片和果实中的蛋白质、RNA、多酚、多糖、单宁、色素等大分子物质以及盐类等小分子杂质.提取的总DNA浓度最高达178 ng/μL,A260/280在1.82~1.94之间,A260/230在1.89~2.04之间.SDS提取法、CTAB提取法和试剂盒提取法所提取的金柑叶片和果实总DNA的扩增效果图谱清晰,多态性高.3种提取方法中,CTAB提取法所提取总DNA浓度较高,纯度最好,可用于后续相关研究.%In order to research effects of different extraction methods on extraction quality of total DNA in Fortunella swingle,taking leaves and fruits in three cultivars of F.swingle as materials,total DNA was extracted by SDS extration method, CTAB extration method and kit extration method, and total DNA concentration, A260/280, A260/230, agarose gel electrophoresis results and SCoT amplified products were compared and analyzed. The results show that the CTAB extraction method and the kit extraction method could better remove protein, RNA, polyphenols, polysaccharide, tannin, pigment and other small molecular substances such as salt and so on in F. swingle leaves and fruits.The total DNA concentration was as high as 178 ng/μL, A260/280was from 1.82 to 1.94, and A260/230was from 1.89 to 2.04. The total DNA in F. swingle leaves and fruits extracted by SDS extration method, CTAB extration method and kit extration method had clear amplification map and high polymorphism.In the three extraction methods,the extracted total DNA by CTAB extraction method had high concentration and the best purity, which could be used for subsequent correlation studies.