摘要
用改良的CTAB法提取不同居群的白木香基因组DNA,通过优化AFLP技术中关键步骤的参数,建立适合白木香的AFLP银染反应体系。结果表明,改良的CTAB法提取的白木香基因组DNA纯度高,完整性好,满足AFLP分析的要求;250ng的基因组DNA用EcoRI和MseI37℃消化3h后可以被完全酶切,酶切产物和接头经16℃连接过夜后,用带有一个选择性碱基的引物和带有3个选择性碱基的引物分别进行预扩增和选择性扩增,扩增产物经变性聚丙烯酰胺凝胶电泳分离,用AgNO3染色得到了清晰的指纹式样,为下一步的遗传分析奠定了基础。
To establish the AFLP silver-stained system suitable for the genetic analysis of Aquilaria sinensis （Lour.） Spreng, the genomic DNA of different A. sinensis populations was extracted by modified CTAB method and an AFLP siliver-stained system suitable for Aquilaria sinensis （Lour.）Spreng was achieved by optimizing the factors in the key steps. The genomic DNA obtained using modified CTAB method has good purity and integrity for AFLP analysis and the fingerprinting of Aquilaria sinensis （Lour.） Spreng achieved with the optimized AFLP silver-stained system was clear. By the modified CTAB method can be obtained the genomic DNA from A. sinensis of different populations with high quality. The optimized AFLP silver-stained system was suitable for the genetic analysis of Aquilaria sinensis （Lour.） Spreng.
出处
《药物生物技术》
CAS
CSCD
2007年第1期14-17,共4页
Pharmaceutical Biotechnology
基金
广东省自然科学基金资助项目(项目编号:06019716)
关键词
白木香
DNA提取
扩增片段长度多态性
银染
建立
Aquilaria sinensis （Lour.） Spreng,DNA extraction, AFLP, Silver-stained, Establishment
