唇螖基因组AFLP反应体系的建立及优化

Establishment and Optimization of AFLP Technological System for Hemibarbus labeo Pallas

以唇螖为实验材料,通过对扩增片段长度多态性(AFLP)的几个关键技术参数进行研究,建立适合于唇螖的AFLP反应体系。结果显示:通过DNA的质量、酶的浓度、酶切时间及扩增时稀释倍数等条件的调整,EcoRI、Tru9I酶切组合和PstI、TaqI酶切组合均能够获得清晰可靠的图谱,二者的结果比较显示,EcoRI、Tru9I酶切组合扩增结果的多态性要稍差于PstI、TaqI酶切组合的扩增结果,为利用AFLP标记对唇螖基因组进行深入研究打下基础。

An AFLP （amplified fragment length polymorphism） analysis system was established with Hemibarbus labeo Pallas by optimizing some factors of the AFLP technical system. The result show that optimizing the quality of template DNA , the concentration of two groups restriction enzymes, the time of the digested reaction and times of pre -amplification products dilution et al can obtained clear and stable amplification patterns. Compared results from two groups of restriction enzymes showed that the group of PstI and TaqI obtained more polymorphie bands than the group of EcoRI and Tru9I. This results provides fundamentals for further molecular studies of using AFLP marker.