摘要
对采自宁波市象山县三门湾沿海滩涂的缢蛏RAPD扩增条件进行了摸索和优化 ,反应体系为 2 5μL时 ,缢蛏的RAPD扩增的最适条件为 94℃预变性 5min ,94℃变性 1min ,37℃退火 1min ,72℃延伸 1min ,进行 4 0个循环 ,然后 72℃总延伸 10min .Taq酶量为 1.0u ,dNTPs为 30 0 μmol/L ,引物浓度为0 .4 μmol/L ,DNA模板浓度为 5 0ng ,Mg2 +浓度为 2 .0mmol/L .对缢蛏DNA进行了初步的遗传分析 。
RAPD(random amplified polymorphic DNA) markers were generated from Sinonovacoula constyricta (Lamarck) of Xiangshan, Ningbo. This paper optimized the reaction factors. Amplifications were performed in the volume of 25 μL of reaction mixture containing buffer(10 mM Tirs HCl pH 8.3, 50 mM KCl), Mg 2+ 2.0 mmol/L,300 μmol/L of total dNTPs, 1.0 u Taq DNA polymerase, 0.4 μmol/L primer, 50 ng DNA template. After an initial heat denaturation at 94 ℃ for 5 min, the reaction mixture was subjected to amplification for 40 cycles consisting of 1 min denaturation at 94 ℃, 1 min at 37 ℃, and 1 min at extension 72 ℃, final extension was 10 min at 72 ℃. Amplified products were separated on 0.8% agarose gel, stained with ethidium bromide and visualized by illumination with ultraviol
出处
《宁波大学学报（理工版）》
CAS
2002年第1期45-49,共5页
Journal of Ningbo University：Natural Science and Engineering Edition
