Related Experiment Video
Updated: Sep 29, 2026

In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA
Published on: May 21, 2015
Checkerboard DNA-DNA hybridisation technology focused on the analysis of Gram-positive cariogenic bacteria
G M Wall-Manning1, C H Sissons, S A Anderson
1Dental Research Group, Department of Pathology and Molecular Medicine, Wellington School of Medicine and Health Sciences, University of Otago, PO Box 7343, Wellington South, New Zealand.
Abstract:
Checkerboard DNA-DNA hybridisation enabled the quantitative analysis of plaque samples against 40 microbial species simultaneously, using digoxygenin-labelled, whole-genome DNA probes. This technique was initially developed to study the predominantly Gram-negative sub-gingival plaque microbiota. The aim of this study was to apply it to a suite of predominantly Gram-positive microorganisms, such as those implicated in cariogenesis. To specifically target Gram-positive species (and Candida albicans) required optimisation and modification of DNA extraction, prehybridisation, hybridisation, and antibody detection conditions. The suitability of the revised technique for clinical and epidemiological studies was confirmed using interproximal plaque from small groups of 5- to 6-year-old children of high (decayed, missing, or filled teeth (dmft)> or =5, n=8) and zero (n=5) caries rates.
Related Concept Videos
Methods of Classification and Identification
Modern Molecular Taxonomy
Southern Blot
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Labeling DNA Probes
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Applications of Molecular Taxonomy
Karyotyping

