南方农业学报2011，Vol.42Issue(3)：233-235,3.
大果油茶基因组DNA提取及ISSR反应体系建立
Isolation of genomic DNA and establishment of ISSR reaction system for Camellia crepnelliana Tutch
摘要
Abstract
[Objective]The present study was undertaken to establish an ISSR-PCR amplification reaction system for Camellia crepnelliana Tutch for providing basis to further research.[Method]The young leaves of Camellia crepnelliana Tutch were used to isolate high-quality genomic DNA using the modified SDS method.The different conditions for ISSR reaction system were optimized.[Result]The extracted genomic DNA was of high quality as revealed by 1.8 to 2.0 absorbance ratio of wavelengths(260/280),did not show any degradation,and was found suitable for ISSR-PCR reaction.The components of the best ISSR-PCR amplification system in a total 25.0 μL reaction volume included 1.0 μL of template DNA (40 ng/μL),2.0 μL of dNTPs (2.5 mmol/L),0.2 μL of Taq DNA polymerase (5 U/μL),1.0 μμL primer (10 μmol/L),2.5 μL 10×PCR buffer(plus Mg2+).The PCR program was set to 5 min at 94℃ for pre-denaturing,followed by 40 cycles of 40 s at 94℃ (denaturation),40 s at 52℃ and 53℃ (annealing)and 90 s at 72℃ (extension),the final extension was set at 72℃ for 7 min.The optimized reaction system gave distinct DNA fingerprinting results.[Conclusion]The established ISSRPCR amplification reaction system may be useful to assess the genetic variability and diversity in Camellia crepnelliana Tutch.关键词
大果油茶/基因组DNA/提取/ISSR反应体系/建立Key words
Camellia crepnelliana Tutch/genomic DNA isolation/ISSR reaction system/genetic diversity分类
农业科技引用本文复制引用
杨翠芳,陈伯伦,黄诚梅,吕维莉..大果油茶基因组DNA提取及ISSR反应体系建立[J].南方农业学报,2011,42(3):233-235,3.基金项目
Guangxi Natural Science Foundation Item(2010GXNSFA0138081) （2010GXNSFA0138081）
