【摘要】 在ＲＡＰＤ反应中，有许多影响结果的稳定性和准确性。本项研究采用浙江省８个板栗主栽品种（无性 系），对ＲＡＰＤ各种反应条件进行了探索，结果表明，板栗理想的反应体系为：２０μｌ反应体积含有１００ ｍｍｏｌ／Ｌ　Ｔｒｉｓ－ＨＣｌ，５００ ｍｍｏｌ／Ｌ ＫＣＬ， ２０ｍｍｏｌ／Ｌ ＭｇＣＬ２， ０．０１％Ｇｅｌａｔｉｎ，ｄＡＴＰ、 ｄＴＴＰ、 ｄＣＴＰ、 ｄＧＴＰ的量各为 １００μｍｏｌ／Ｌ， ５０－２００ｎｇ引物， ０．７５－１．０单位Ｔａｑ酶， ２－１０ｎｇ模板ＤＮＡ。反应条件为： ９４℃预变性２ｍｉｎ， 再９４℃　３０ｓ、４０℃３０ｓ、７２℃１．５ｍｉｎ扩增３８个循环；最后在７２℃延伸７ｍｉｎ。应用上述反应体系进行板栗 ＲＡＰＤ反应，扩增产物在１．０％琼脂糖凝胶中电泳，经ＥＢ染色后在紫外灯下观察照相，可获得满意的ＤＮＡ 指纹图谱。更多还原

【Abstract】 Many factors influence stability and accuracy of the result in RAPD reaction. Reaction conditions of RAPD on 8 main cultivated Castenea mollissima clones in Zhejiang province showed the ideal reaction condition: 100 m mol/L This-HCI, 500 m mol/L KCL, 20 m mol/L MgCl2, 0.01% Gelatin, 100 μ mol/L dATP, dTTP, dCTP and dGTP in 20 μ l reaction content with 50-200 ng primer, 0.75-l.0 unit Taq enzyme and 2-10 ng template DNA; reaction condition: 2 minutes Pre-denaturation at 94℃, enlarging 38 circulation at 94℃ in 30 seconds, at 40℃ in 30 seconds and at 72℃ in 1.5 minute; at last, elongation at 72℃ in 7 minutes. RAPD reaction of Castanca mollissima with above-mentioned reaction system, and electrophoresis in l.0% agar-gel of enlarged material, observing and taking photos under ultraviolet lamp after EB staining, satisfied DNA finger print could be obtained.更多还原