摘要
以充分展开的山核桃Carya cathayensis嫩叶为材料,利用改良CTAB(十六烷基三甲基溴化铵)法与CTAB裂解-硅珠吸附法提取山核桃总DNA,对适合于山核桃叶片的AFLP(扩增片段长度多态性)分析体系进行了摸索。结果表明:"!改良CTAB法适用于从山核桃叶中提取高质量的DNA,提取的DNA可用于AFLP的分析;#"采用EcoR I/MseⅠ酶切组合,酶切连接一步法与两步法的效果没有明显的差异,但一步法操作更为简便;&%适合于山核桃叶AFLP分析的引物组合为E-ACT+M-CAT,E-ACT+M-CTG,E-ACG+M-CAT,E-ACG+M-CTG。扩增产物经60 g.L-1变性聚丙烯酰胺凝胶电泳后进行银染,条带信号强度一致性好,条带分布均匀、清晰,分辨率较高。
Total DNA was extracted from young, fully opened leaves of Carya cathayensis (hickory) by an improved cetyl trimethylammonium bromide (CTAB) method and a CTAB-silicon method, based on which an amplified fragment length polymorphism (AFLP)analysis system for hickory was developed. An electrophoresis was used on the amplified products in a 6% denatured polyacrylamide gel, followed by silver staining, with clearly identified bands shown. Specifically: (1)the improved CTAB method was appropriate for DNA extraction from hickory leaves with extracted DNA qualified for AFLP analysis; (2)with a combination of EcoR I and Mse I for enzyme cutting, the results were the same for the one-step method combining enzyme cutting and adaptor ligation and the two-step method which separated them with the one-step method being much easier; and (3)there were four primer pairs suitable for this AFLP analysis, namely E-ACT + M-CAT, E-ACG + M-CAT, E-ACG + M-CAT, and E-ACG + M-ATG.
出处
《浙江林学院学报》
CAS
CSCD
北大核心
2008年第4期532-537,共6页
Journal of Zhejiang Forestry College
基金
浙江省自然科学基金资助项目(Y304416)
浙江省现代森林培育技术重点实验室开放基金资助项目(200503)
国家自然科学基金资助项目(30771752)
关键词
植物学
山核桃
DNA提取
扩增片段长度多态性(AFLP)
botany
Carya cathayensis (hickory)
extraction of DNA
amplified fragment length poly morphism (AFLP)