Olsen A S, Combs J, Garcia E, Elliott J, Amemiya C, de Jong P, Threadgill G
Lawrence Livermore National Laboratory, Livermore, CA 94550.
Biotechniques. 1993 Jan;14(1):116-7, 120-3.
We report here on a system for automated preparation of high-density colony filters of arrayed libraries using the high density replicating system (HDR) for the Beckman Biomek 1000 robotic workstation. This system, consisting of a 96-pin tool, a sterilization station and controlling software, transfers samples from microplates onto target membranes in arrays up to 36 times the density of a 96-well microplate. The transfer operation can be completely automated with the addition of the Biomek Side Loader System, which consists of a robotic arm capable of transferring plates and filters between the Biomek working tablet and a storage area. Using the complete system, we are able to plate 32 replica filters (8 x 12 cm), each containing the clones from 16 different microplates (i.e., 1536 clones per filter), in a 16-h overnight run without any operator intervention. We describe conditions used for transfer of bacterial yeast colonies and fixation of DNA to the membranes, and we illustrate hybridization results obtained with cosmid and YAC filters.
我们在此报告一种利用贝克曼Biomek 1000机器人工作站的高密度复制系统（HDR）自动制备阵列文库高密度菌落滤膜的系统。该系统由一个96针工具、一个消毒站和控制软件组成，可将样品从微孔板转移到密度高达96孔微孔板36倍的阵列靶膜上。添加Biomek侧装载系统后，转移操作可完全自动化，该系统包括一个能够在Biomek工作平板和存储区域之间转移平板和滤膜的机械臂。使用完整的系统，我们能够在无需任何操作员干预的情况下，在16小时的过夜运行中铺板32个复制滤膜（8×12厘米），每个滤膜包含来自16个不同微孔板的克隆（即每个滤膜1536个克隆）。我们描述了用于转移细菌酵母菌落和将DNA固定到膜上的条件，并展示了用黏粒和酵母人工染色体滤膜获得的杂交结果。
