Method combining BAC film and positive staining for the characterization of DNA intermediates by dark-field electron microscopy.

ABSTRACT: DNA intermediate structures are formed in all major pathways of DNA metabolism. Transmission electron microscopy (TEM) is a tool of choice to study their choreography and has led to major advances in the understanding of these mechanisms, particularly those of homologous recombination (HR) and replication. In this article, we describe specific TEM procedures dedicated to the structural characterization of DNA intermediates formed during these processes. These particular DNA species contain single-stranded DNA regions and/or branched structures, which require controlling both the DNA molecules spreading and their staining for subsequent visualization using dark-field imaging mode. Combining BAC (benzyl dimethyl alkyl ammonium chloride) film hyperphase with positive staining and dark-field T

SUBMITTER: Benureau Y 

PROVIDER: S-EPMC7474861 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature