Gergen J P, Stern R H, Wensink P C
Nucleic Acids Res. 1979 Dec 20;7(8):2115-36. doi: 10.1093/nar/7.8.2115.
A permanent, ordered collection of 23,000 recombinant DNA plasmids containing Drosophila melanogaster DNA has been established. Simple and practical methods for storing and manipulating this collection were developed. In addition, an improved, simple and inexpensive method for making paper filter replicas of such an ordered collection and of a high density (10,000 colonies/petri dish) unordered collection was developed. These filter replicas are suitable for nucleic acid hybridization screens of recombinant DNA colinies and each filter replica can be used for many (greater than 5) successive screens. The kinetics of this hybridization reaction were examined and allow design of experiments that detect colony complementarity to a nucleic acid that is 0.5% of the hybridization probe.
已建立了一个包含23,000个含有黑腹果蝇DNA的重组DNA质粒的永久性、有序文库。开发了用于存储和操作该文库的简单实用方法。此外，还开发了一种改进的、简单且廉价的方法，用于制作这种有序文库以及高密度（每平皿10,000个菌落）无序文库的滤纸复制品。这些滤纸复制品适用于重组DNA菌落的核酸杂交筛选，并且每个滤纸复制品可用于多次（大于5次）连续筛选。研究了这种杂交反应的动力学，并据此设计实验，以检测与杂交探针0.5%的核酸具有菌落互补性的情况。
