Labeling DNA for single-molecule experiments: methods of labeling internal specific sequences on double-stranded DNA
Authors:
Open Access
- 6 July 2011
- journal article
- review article
- Published by Royal Society of Chemistry (RSC) in Nanoscale
- Vol. 3 (8) , 3027-3039
Abstract
This review is a practical guide for experimentalists interested in specifically labeling internal sequences on double-stranded (ds) DNA molecules for single-molecule experiments. We describe six labeling approaches demonstrated in a single-molecule context and discuss the merits and drawbacks of each approach with particular attention to the amount of specialized training and reagents required. By evaluating each approach according to criteria relevant to single-molecule experiments, including labeling yield and compatibility with cofactors such as Mg2+, we provide a simple reference for selecting a labeling method for given experimental constraints. Intended for non-specialists seeking accessible solutions to DNA labeling challenges, the approaches outlined emphasize simplicity, robustness, suitability for use by non-biologists, and utility in diverse single-molecule experiments.Keywords
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