摘要
为获得桉树种质鉴定、辅助育种所需稳定的分子标记,本研究以生产中常用的24个桉树无性系基因组DNA为材料,对AFLP的反应体系和参数进行优化,发现高纯度的模板最低用量为100 ng;EcoRⅠ、MseⅠ用量不低于0.5 U;酶切连接时间需要大于4 h,但不多于14 h。预扩增获得50～700 bp产物;该产物稀释5倍后用于选择性扩增。聚丙烯酰胺胶电泳检测温度以20℃为佳。
In order to get stability molecular markers for Euca/yptus gennplasm and molecular - marker assist- ed breeding, 24 Euca/yptus clones commonly used in plantations were selected as the test materials for genom- ic DNA extraction. By optimizing each reztion procedure, an AFLP reaction system was established. This system included 100 ng high purity genome DNA and no less than 0.5 U EcoR I and Mse I respectively, 4- 14 hour enzyme digestion and DNA reunion, 5 times diluted pre - amplification products for selective amplifica- tion. The fragments were 50 bp to 700 bp in the pre - amplification products, lastly, the best temperature for ~lvacrvlamide gel electrophoresis was 20 ℃.
出处
《桉树科技》
2013年第1期20-24,共5页
Eucalypt Science & Technology
基金
广西青年基金项目"不同生化类型互叶白千层分子生物学鉴定"(2013GXNSFBA019094)
林业公益性行业科研专项"桉树生态经营及产业升级关键技术研究"(201104003)
广西科学研究与技术开发计划课题"高产优质多抗桉树新品种选育"(桂科攻1123004-3A)
关键词
桉树
AFLP
体系优化
Eucalyptus
AFLP
reaction system optimization
