北京林业大学学报2012，Vol.34Issue(1)：51-57,7.
基于毛细管电泳的柳树AFLP分子标记研究
AFLP molecular marker of Salix spp.based on apillary electrophoresis
摘要
Abstract
In this paper,we established and optimized the AFLP molecular marker technology of Salix spp.based on capillary electrophoresis as well as simplify analysis flow of AFLP in order to construct Salix spp.linkage mapping and the implement of molecular breeding.First,the high-quality genomic DNA was extracted for AFLP analysis.Then the genomic DNA was completely digested by restriction enzyme and ligased with oligonucleotide adapters successfully.We got the products via pre-amplification and selective amplification.At last,the selective amplification products were analyzed by capillary electrophoresis.Results showed that improved CTAB method was selected to extract genomic DNA,450 ng of genomic DNA was used in digestion with EcoRⅠ and MseⅠ for 2 hours respectively,and ligased overnight.In selective amplification,0.3 mmol/L dNTP,1.5 mmol/L Mg2＋,0.125 μmol/L primers,0.025 U/μL polymerase and 20 times dilution of pre-amplification products were desirable.Through many repeats,this system has been proved to be feasible in Salix spp.AFLP-capillary electrophoresis analysis.关键词
柳树/AFLP/毛细管电泳Key words
Salix spp./AFLP/capillary electrophoresis分类
农业科技引用本文复制引用
吕金辉,胡建军,卢孟柱..基于毛细管电泳的柳树AFLP分子标记研究[J].北京林业大学学报,2012,34(1):51-57,7.基金项目
“863”国家高技术研究发展计划项目 （）
转基因生物新品种培育科技重大专项 （）
