Ramanujam P, Fogerty S, Heiser W, Jolly J
Pharmacia P-L Biochemicals, Inc., Milwaukee, WI 53202.
Biotechniques. 1990 May;8(5):556-63.
A rapid method for electrophoresis of DNA-protein complexes is described. Popular "gel-shift" assays are performed using Pharmacia PhastSystem with its convenience of pre-cast gels and buffer strips and microprocessor-controlled high-resolution separation. Using this system, the products of a DNA binding reaction (DNA-protein complexes) can be separated from "free" DNA in less than one hour. DNA fragments as well as oligonucleotides have been used as targets with partially purified extracts containing sequence-specific DNA binding proteins. We present here a comparison of results of gel-shift assays obtained by conventional techniques and by our rapid "PhastShift" method which reduces the time, effort and technical expertise necessary to obtain reproducible results.
本文描述了一种用于DNA-蛋白质复合物电泳的快速方法。常用的“凝胶迁移”分析是使用Pharmacia PhastSystem进行的,它具有预制凝胶、缓冲条带以及微处理器控制的高分辨率分离等便利之处。使用该系统,DNA结合反应的产物(DNA-蛋白质复合物)能够在不到一小时的时间内与“游离”DNA分离。DNA片段以及寡核苷酸已被用作靶标,与含有序列特异性DNA结合蛋白的部分纯化提取物进行反应。我们在此展示了通过传统技术和我们的快速“PhastShift”方法获得的凝胶迁移分析结果的比较,后者减少了获得可重复结果所需的时间、精力和技术专业知识。