摘要
以克氏原螯虾DNA为材料,对AFLP分析中的基因组酶切时间、选择性扩增中预扩增产物稀释倍数、Mg2+浓度等3个因素进行了比较。结果表明,基因组用EcoRⅠ和MseⅠ双酶切5 h,选择性扩增的PCR反应体系中Mg2+1.5 mmol/L是进行克氏原螯虾AFLP分析的最佳反应条件,能够得到丰富稳定的带纹。而预扩增产物稀释倍数对选择性扩增没有明显影响。该体系的构建为AFLP技术在克氏原螯虾相关研究中的应用奠定了基础。
By using genomic DNA of Procambarus clorkii, the time for DNA double enzymes digestion reaction,the dilution multiple for the pre-amplification production and the concentration of Mg2＋ in the selective amplification reactions system were analyzed. The results showed using 5h for double enzymes digestion and Mg2. 1.5 mmol/L were the best for AFLP analysis of P. clarkii. And no change was found when adjusting the dilution multiple for the pre-amplification production. The establishment of the system might lay a foundation for the application of AFLP techniques to the relative research of P. clarkii.
出处
《湖北农业科学》
北大核心
2009年第11期2804-2807,共4页
Hubei Agricultural Sciences
基金
国家科技支撑计划项目(2007BAD37B01)
