摘要
目的:建立一个适于日本沼虾研究用的AFLP反应体系。方法:以日本沼虾DNA为材料,对基因组酶切时间、选择性扩增中Mg2+、dNTP浓度、预扩增产物稀释倍数及选扩性引物M+3/E+3配比等进行了比较分析。结果:酶切5h,选扩25μl PCR反应体系中Mg2+2mmol/L,dNTP 1.2mmol/L,预扩产物稀释40倍,选扩引物M+3/E+3配比为8∶1,所得产物在毛细管电泳中可得到稳定的结果。结论:该体系的构建为AFLP技术在日本沼虾相关研究中的应用奠定了基础。
Objective：To establish an appropriate AFLP（Amplified Fragment Length Polymorphism）analysis system for Macrobrachium nipponensis. Methods： By using genomic DNA of M. niponensis adjusting the time for DNA double enzymes digestion reaction, the dilute multiple for the preamplification production and the concentration of Mg^2 ＋ , dNTP in the selective amplification reactions system, different proportion in selective primers M ＋ 3/E ＋ 3 to select the optimization. Results. It was proved that quite steady results could be achieved in capillarity electrophoresis by using 5h for double enzymes digestion, Mg2; 2mmol/L , dNTP 1.2mmol/L , 40 dilute multiple for the preamplification production, 8：1 for M ＋ 3/E ＋ 3 selective primer in 25 μl PCR reaction volume. Conclusion：The establishment of the system might lay a foundation for the application of AFLP techniques to the relative research of M. nipponensis.
出处
《生物技术》
CAS
CSCD
2008年第1期36-39,共4页
Biotechnology
基金
国家"十一五"科技支撑计划项目(2006BAD01A13
2006BAD03B07-02)
2007年度农业科技跨越计划项目
江苏省高技术研究项目(BG2007328)资助
关键词
日本沼虾
AFLP
反应体系
Macrobrachium nipponensis
AFLP
reaction system
