东方百合杂种后代试管苗叶片总DNA提取方法的选择  

Choosing Suitable Method to Extract Genomic DNA from the Leaves of In-tube Seedling of the Hybrid Offspring of Oriental Lily

作　　者：胡凤荣[1,2] 国荣荣[2] 王斐[2] 王江勇[3] 

机构地区：[1]北京林业大学,北京100083 [2]南京林业大学,南京210037 [3]山东省果树研究所,泰安271000

出　　处：《分子植物育种》2013年第2期262-266,共5页Molecular Plant Breeding

基　　金：江苏省高校优势学科建设工程项目资助

摘　　要：以东方百合‘Sorbonne’בFrancia’的杂种后代试管苗叶片为材料,采用CTAB-高盐法、简易CTAB法、CTAB-硅珠法和SDS-CTAB法等4种DNA提取方法,比较DNA耗时、纯度、得率、浓度和质量等指标,以期为东方百合‘Sorbonne’בFrancia’杂种群体大量杂种后代试管苗叶片总DNA的提取筛选适宜方法。结果表明,除CTAB-硅珠法未获得DNA,其余3种方法均获得条带清晰、完整性好的DNA。在耗时方面,CTAB-硅珠法<简易CTAB法<CTAB-高盐法<SDS-CTAB法;在纯度方面,简易CTAB法纯度高于SDS-CTAB法、CTAB-高盐法,质量最佳,OD260/OD280为1.98,OD260/OD230为2.01,浓度为190.01ng/μL,得率为57.01μg/g。综合比较,采用简易CTAB法进行杂种群体大量试管苗叶片总DNA提取效果更佳。In order to screen the ideal method to extract genomic DNA from the Leaves of in-tube seedling of oriental lily, the hybrids offspring derived from the cross of Sorbonne × Francia were used as the experimental materials in this study. We compared the DAN quality and extracting efficiency of the employed DNA extraction methods, such as SDS-CTAB, Simple and ease CTAB, CTAB-high salt precipitation and CTAB-silica purification, in the facets of the purity, yield, concentration, and time consuming to choosing the ideal DNA extraction method. Three methods can be used to extract DNA with good banding profiles and Integrity except CTAB-silica purification missing DNA. The orders of the time consuming were from more to less following as CTAB-silica purification, Simple and ease CTAB, CTAB-high salt precipitation and SDS-CTAB. In the term of DNA purity, the simple CTAB method got the highest DNA purity than that of the SDS-CTAB and CTAB-high salt precipitation, which the extracted DNA with the excellent quality with OD260/OD280, 1.98, OD260/ODz0, 2.01; while the concentration of the extracted DNA reached 190.01 ng/μ.L as well as 57.01 μg/g of the DNA yield. Above all, Simple and ease CTAB is the most suitable way to extract a great quantity of genomic DNA from the leaves of in-tube seedling of the hybrid offspring of oriental lily.

关 键 词：东方百合 杂种后代 试管苗叶片 基因组DNA 提取方法 

分 类 号：S682.29[农业科学—观赏园艺]