摘要
为从珍珠黄杨幼叶中提取高质量的总DNA,比较了1%CTAB、2%CTAB、4%CTAB和1.5%SDS4种DNA提取方法。结果表明:2%CTAB提取的DNAA260/A280值最好,ISSR-PCR扩增效果最佳,是有效提取珍珠黄杨基因组DNA的方法。并在此基础上,以(CT)8G为引物,采用单因素实验法,确立了适合珍珠黄杨的ISSR-PCR反应体系:在总体积20μL的反应体系中,含30ngDNA模板,2.0mmol/LMg2+,0.30μmol/L引物,0.20mmol/LdNTPs,1UTaq酶。
In this paper, four different DNA-extracting methods （1% CTAB. 2% CTAB. 4% CTAB and 1.5% SDS） were employed to be compared in order to isolate high-quality genomic DNA from young leaves of Buxus sinic vat. parvifolia. The results showed that the method with 2% CTAB was the best one, which the extracted DNA had an ideal A260/A280 ratio and nice banding pattern amplified by ISSR-PCR. We could recommend the ISSR- PCR reaction system for Buxus sinic var. parvifolia based on optimization with primer （CT）8G by the single factor experiment. The components of 20 μL reaction volume would be as follows： 30 ng genomic DNA as template, 2.0 mmol/L Mg^2＋, 0.30μmol/L primer, 0.20 mmol/L dNTPs, 1 U Taq polymerase and added ddH20 to total volume.
出处
《分子植物育种》
CAS
CSCD
2007年第1期145-149,150,共6页
Molecular Plant Breeding
基金
国家十五科技攻关项目(2004BA515B0102)资助。
