桃属植物随机扩增多态性DNA分析的最适反应条件

Factors influencing RAPD band patterns in Prunus Persica

以Ｙｕｕｚｏｒａ 桃为试材对桃属植物随机扩增多态性ＤＮＡ（ＲＡＰＤ） 分析的最佳反应条件进行了研究．结果表明桃属植物ＲＡＰＤ分析的最佳反应条件为：总反应混合溶液５ｕＬ 时，添加模板ＤＮＡ１０ ～３０ ｎｇ，Ｔａｑ ＤＮＡ 聚合酶１ ～１ ．５ 个单位，随机引物１０ ～２０ ｐｍｏｌ，ｄＮＴＰ１００ ～２００ ｕＭ，ＭｇＣｌ２ ２ ．０ ～３ ．０ ｍ Ｍ 及５ ｕＬ１０ 倍反应缓冲液条件下扩增５０ ～５５ 个循环，能够获得具有良好的稳定性和重复性的ＤＮＡ扩增产物．ＰＣＲ反应溶液的各组成因素如低于最适浓度范围时会导致扩增带数的减少甚至无带，如高于最适反应浓度范围，则会出现带的弥散或缺失．

This study was carried out to investigate several factors affecting RAPD band patterns in Prunus Persica and to establish the optimum RAPD conditions for Prunus Persica .According to the results the optimized RAPD reaction conditions for Prunus persica were 5～30 ng template DNA,10～20 pmol random primer,100～200 uM dNTP,2.0～3.0 mM MgCl 2,1.5～2.0 unit Taq DNA polymerase in a total of 50uL reaction mixture,and 50～55 cycles of amplification produced best RAPD band patterns.Under optimized reaction conditions,RAPD band patterns of Prunus persica were quite repeatable.Lower concentration caused the decrement or lack of RAPD bands and higher amount resulted in smear of RAPD bands or failure of amplification.