ramp在提高桂圆RAPD分析效率中的应用研究

Research on Application of Ramp to Improve the Efficiency of RAPD in Longan

[目的]检验一种提高RAPD技术效率的新方法（调整ramp）。[方法]以四川、福建、广西、广东和海南的桂圆为材料,用改进的CTAB法提取桂圆叶的总DNA,选取了18种重复性好的随机引物来进行RAPD扩增。在RAPD试验中,参数ramp分别设为0.3和3.0℃/s,比较它们对桂圆样本RAPD扩增效果的影响,并对几个产地桂圆的亲缘关系进行分析。[结果]当ramp由3.0℃/s降低为0.3℃/s时,可明显提高RAPD的分辨率和产量;通过聚类分析表明5个品种桂圆的遗传距离为0.69～0.76。[结论]该试验证明降低ramp速度可明显提高RAPD技术的分辨率和产量,为RAPD技术提供了一种新的方法。

[Objective] This study aimed to detect a modified method for improving the efficiency of RAPD technology.[Method] Longans collected from Sichuan,Fujian,Guangxi,Guangdong and Hainan were used as experimental materials.Total genomic DNA was extracted from longan leaves with modified CTAB method.18 random primers were selected for RAPD amplification with ramp rates of 0.3 ℃/s and 3.0 ℃/s,respectively.Effects of different ramp rates on RAPD amplification were compared and the relationship among longan samples was analyzed.[Result] The resolution and yield could be improved significantly by declining the ramp rate from 3.0 ℃/s to 0.3 ℃/s.Cluster analysis showed that similarity coefficient of the five varieties of longan ranged from 0.69 to 0.76.[Conclusion] This study had confirmed that reducing ramp rate could significantly increase the resolution and yield of RAPD technology,which had provided a new method to improve the efficiency of RAPD.