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以云南马铃薯主要栽培品种为材料 ,使用进口PCR仪 ,对马铃薯RAPD分析中的DNA模板浓度、引物浓度、dNTP浓度以及PCR扩增反应循环次数进行了研究。结果表明适用于马铃薯RAPD反应的最佳条件为 :模板DNA1 5～ 5 0ng ,1 0 mer随机引物 2 5～ 5 0ng ,dNTP2 0 0 μM ,扩增反应周期 40～ 45个循环。每个循环包括 94℃变性 1min ,37℃退火 1min ,72℃延伸 2min。使用这一RAPD程序对云南的主要栽培品种进行品种鉴别 ,能有效区分出所有参试品种 ,检测出品种CFK6 9 1遗传背景特殊 ,较之于普通栽培品种亲缘关系较远。本实验建立了适宜马铃薯RAPD分析的PCR程序及条件 ,为RAPD应用于马铃薯的遗传研究打下了基础。
Taking the main potato cultivars from Yunnan as materials, the PCR instrument was used to study the DNA template concentration, primer concentration, dNTP concentration and PCR amplification reaction cycle number in potato RAPD analysis. The results showed that the optimum conditions for potato RAPD reaction were as follows: template DNA 1.5 ~ 5 0ng, 10 ~ random primer 25 ~ 50ng, dNTP2 0 0 μM, amplification reaction cycle 40 ~ 45 cycles. Each cycle includes denaturation at 94 ° C for 1 min, annealing at 37 ° C for 1 min, extension at 72 ° C for 2 min. Using this RAPD program to identify the main cultivars in Yunnan Province, all the tested cultivars can be effectively distinguished, and the genetic background of CFK6 9 1 was detected to be more distantly related to that of ordinary cultivars. In this study, we established a PCR program and conditions suitable for RAPD analysis of potato, which laid the foundation for the genetic research of RAPD in potato.