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用 4 5条随机引物对不同来源的 9株嗜水气单胞菌进行了 DNA指纹分析 ,其中 10条引物呈现良好的多态性和稳定性。建立的随机扩增多态 DNA(RAPD)反应的最佳条件 :模板 DNA 2 0 ng,随机引物 0 .8μm ol/ L ,d NTP 15 0μmol/ L,Mg2 + 3.5 mm ol/ L,Taq酶 2 .0 U;95℃预变性 5 min,94℃变性 6 0 s,36℃退火 70 s,72℃延伸 12 0 s,4 0个循环。初步建立了嗜水气单胞菌的 RAPD指纹图谱 ,分析了 9株嗜水气单胞菌之间的遗传距离。据此将 9株菌归为 3个组 ,为该菌的分型提供了新的方法。
DNA fingerprinting was performed on 9 strains of Aeromonas hydrophila from different sources using 45 random primers. Among them, 10 primers showed good polymorphism and stability. The optimal conditions for the RAPD reaction were as follows: template DNA 20 ng, random primer 0.8 μmol / L, dNTP 15 0 μmol / L, Mg 2+ 3.5 mm ol / L, Taq enzyme 2 .0 U; denaturation at 95 ° C for 5 min, denaturation at 94 ° C for 60 s, annealing at 36 ° C for 70 s, extension at 72 ° C for 12 s, 40 cycles. RAPD fingerprinting of Aeromonas hydrophila was preliminarily established, and the genetic distance between 9 strains of Aeromonas hydrophila was analyzed. According to this, 9 strains were classified into 3 groups, which provided a new method for the typing of this strain.