摘要
以古老月季品种‘玉玲珑’为试材,利用CTAB法对月季进行总DNA提取,对其PCR扩增条件中的主要因素进行了多梯度的优化,总反应体系为25μL,其中引物浓度为0.4μmol/L、Mg2+浓度为2.0 mmol/L、dNTP为0.2 mmol/L、Taq酶用量为1.0 U、DNA模板用量为70 ng。利用该优化体系对部分月季品种和野生种进行PCR扩增和电泳检测,扩增条带清晰,结果稳定,测序结果理想,表明该体系适用于月季的ITS序列分析。
DNA of rose Yulinglong was extracted with CTAB. Principal factors of PCR had different concentrations and their variation changed the result of ITS -PCR. Factors affecting the ITS results of rose were studied and the best reaction system of ITS -PCR was： 0. 4 umol/L of each primer, 2. 0 mmol/L Mg^2＋, 0. 2 mmol/L of dNTPs, 1.0 U of Taq DNA polymerase and 70 ng template DNA in 25 μL reaction system. Using above PCR system, ITS fragments of some rose cultivars and species were obtained. The clearity and stability of amplification indicated this system was suitable for analyzing ITS sequences in roses.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2009年第6期756-758,共3页
Journal of Huazhong Agricultural University
基金
国家自然科学基金项目(30660117)
国家高技术研究发展计划项目(2006AA100109)
云南省科技计划项目(2006NG14)资助
关键词
ITS
月季
优化
ITS
rose
optimization
