Semikozova O P, Berberov E, Shaĭkhaev G O, Sulimova G E
Mol Gen Mikrobiol Virusol. 1992 Nov-Dec(11-12):14-9.
A set of experiments has been conducted to choose the optimal conditions for DNA transfer and fixation on two types of the nitrocellulose, three types of nylon membranes and on capron filters. The buffer capillary transfer systems, electroblotting and gel hybridization are analyzed. Two techniques for DNA binding have been tested under different transfer conditions for all the membrane types: a vacuum fixation at 80 degrees C and a UV-exposure. The results indicate the critical dependence of the efficiency of blot-hybridization on the conditions of UV-treatment. The UV-exposure longer or shorter than the optimal one resulted in a loss of the hybridization efficiency. The optimal DNA-transfer and fixation conditions are recommended for all the membranes tested. The dependence of the optimal transfer and binding conditions on the specific characteristics of different membrane types was demonstrated for maximal sensitivity of the blot-hybridization.
已经进行了一系列实验,以选择在两种类型的硝酸纤维素、三种类型的尼龙膜和卡普纶滤膜上进行DNA转移和固定的最佳条件。分析了缓冲液毛细管转移系统、电印迹和凝胶杂交。在所有膜类型的不同转移条件下测试了两种DNA结合技术:80摄氏度真空固定和紫外线照射。结果表明,印迹杂交效率对紫外线处理条件有至关重要的依赖性。紫外线照射时间长于或短于最佳时间都会导致杂交效率损失。针对所有测试的膜推荐了最佳的DNA转移和固定条件。为了使印迹杂交具有最大灵敏度,证明了最佳转移和结合条件对不同膜类型特定特性的依赖性。