Filter replicas and permanent collections of recombinant DNA plasmids

作者：J. Peter Gergen, Ralph H. Stern, Pieter C. Wensink · 发表于：Nucleic Acids Research · 年份：1979 · DOI：10.1093/nar/7.8.2115 · 被引用次数：355 · 研究领域：Advanced biosensing and bioanalysis techniques、DNA and Biological Computing、Bacteriophages and microbial interactions

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.