正交试验法优化芍药基因组DNA CTAB法提取工艺.pdfVIP

安徽农业科学，Journal ofAnhuiA ．Sei．2008，36(15)：6190—6191 责任编辑 陈玉敏 责任校对 张敏 正交试验法优化芍药基因组DNA CTAB法提取工艺 段国锋，刘慧(山西农业大学园艺学院，山西太谷030801) 摘要 [目的]为芍药的杂交育种提供试验依据。[方法]通过正交试验，对利用芍药基因组DNA的CATB法提取工艺进行优化。[结果] CTAB法提取DNA的0D ／0D， 为1．8～1．9，不同处理间差异较明显。除泳道c7电泳结果较差外，其他泳道DNA的完整性都相对较 好。除c1、c2泳道RNA拖尾现象较严重外，c3、c4、c5c6泳道脱尾现象均较轻微，而c8和c9电泳条带较为整齐。CTAB法提取芍药基因 组DNA的最佳配方为：CrAB 0．75 g、NaC1 3 EDT_A 2．5 ml、Tris—HC1 5 ml，pH值8．0。[结论]利用该CATB方法，可获得高纯度芍药DNA。 关键词 芍药；基因组总DNA；正交试验 中图分类号 $664 文献标识码 A 文章编号 0517—6611(2008)15—06190—02 Orthogonal Test Oil the Optimization ofExtraction Process of Genomic DNA from Paeonia lactiflom Pal1．bY Using CATB Method DUAN Guo-~eng et al (College of Horticulture，Shanxi Agricultural University，Taigu，Shar~xi 030801) Abstract 『Objective]The aim ofthe research was to provide the experimental basis，for cross breeding of Paeonia lactiflora Pal1．．[Method J In the or— thogonal test，the extraction process ofthe genomie DNA from P．1actifiora was optimized．[Result]卟e 0D260／O value of the extracted DNA by CTAB method was 1．8～1．9 and there were inore obvious difference among different treatments．Except the eleetmphoresis resulis of gelwork lane were worse，the integrality of DNA in other gelwork lanes were all relatively better．Except the tailing pl1en伽 r诅of cl，c2 gelwork lanes were inore serious， the tailing phenomena of c3．c4．c5 and c6 gelwork lanes were all inore slight and the electrophoretie bands of c8 and c9． best fomula of extracting the genomic DNA from P．1actifiora by CATB method was CTAB of0．75 g，NaC1 of3 g，EDTA of2．5 m1， s—HC1 of 5 m1，pH value of 8．0．1Con— elusion]DNA of P．1actiflora 出high pmity could be obtained by using this C加 method． Key words Paeonia lactiflora Pal1．；Total genomie DNA；Orthogonal experiment 中国芍药作为观赏植物，已有3 900多年的栽培历史，已 光光度计下测定260和280 nm处的吸收值，计算 0D26o／ 被广泛应用于医药、保健、饮料、食品、香精等领域。对于芍 OD2so值，并利用琼脂糖凝胶电泳进行质量完整性的检测。 药品种的分类，国内外专家提出了许多方案l1 J。这些方案主