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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。
There are many stability and accuracy effects in the RAPD reaction. In this study, 8 chestnut cultivars (clones) from Zhejiang province were used to investigate the reaction conditions of RAPD. The results showed that the ideal reaction system of chestnut was as follows: 20 μl reaction volume containing 100 mmol / L Tris-HCl, 500 100 mmol / L KCL, 20 mmol / L MgCL2, 0.01% Gelatin, dATP, dTTP, dCTP and dGTP, 50-200 ng primers, 0.75-1.0 Taq enzyme, Template DNA. The reaction conditions were as follows: pre-denaturation at 94 ° C for 2 min, amplification for 38 cycles at 94 ° C for 30 s, 40 ° C for 30 s, and 72 ° C for 1.5 min; and finally at 72 ° C for 7 min. Chestnut RAPD reaction was carried out by using the above reaction system. The amplified product was electrophoresed on a 1.0% agarose gel and stained with EB under ultraviolet light to obtain satisfactory DNA fingerprinting.