正交试验法优化芍药基因组DNA CTAB法提取工艺

Orthogonal Test on the Optimization of Extraction Process of Genomic DNA from Paeonia lactiflora Pall.by Using CATB Method

[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OD260/OD280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c1c、2泳道RNA拖尾现象较严重外,c3、c4、c5c、6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：CTAB 0.75 g、NaCl 3 g、EDTA 2.5 mlT、ris-HCl 5 ml,pH值8.0。[结论]利用该CATB方法,可获得高纯度芍药DNA。

[Objective] The aim of the research was to provide the experimental basis for cross breeding of Paeonia lactiflora Pall..[Method] In the orthogonal test,the extraction process of the genomic DNA from P.lactiflora was optimized.[Result] The OD260/OD280 value of the extracted DNA by CTAB method was 1.8～1.9 and there were more obvious difference among different treatments.Except the electrophoresis results of gelwork lane were worse,the integrality of DNA in other gelwork lanes were all relatively better.Except the tailing phenomena of c1,c2 gelwork lanes were more serious,the tailing phenomena of c3,c4,c5 and c6 gelwork lanes were all more slight and the electrophoretic bands of c8 and c9.The best fomula of extracting the genomic DNA from P.lactiflora by CATB method was CTAB of 0.75 g,NaCl of 3 g,EDTA of 2.5 ml,Tris-HCl of 5 ml,pH value of 8.0.[Conclusion] DNA of P.lactiflora with high purity could be obtained by using this CATB method.