Detection of microbial DNA sequences by colony hybridization

Abstract

Colony hybridization, the technique of lysis in situ of bacterial colonies on filters followed by denaturation and binding of the cellular DNA, allowing hybridization to DNA or RNA probes, was first reported in 1975 [3]. It was developed initially for screening Escherichia coli transformants in cloning experiments, to identify colonies containing a specific gene. Subsequently many variations of the method have been generated, appropriate for different applications, including analysis of environmental isolates. The main benefit of in situ colony hybridization is that isolates containing a target sequence can be identified from amongst hundreds or thousands of colonies and recovered for further analysis from either the original plate or a replicate.