Propidium iodide and PicoGreen as dyes for the DNA fluorescence correlation spectroscopy measurements.
Many experimental designs, in which nucleic acid conformational changes are of interest, require reliable fluorescence labeling. The appropriate fluorescence probe should have suitable optical properties and, more importantly, should not interfere with the investigated processes. In order to avoid chemical modifications the fluorescence label needs to be associated with nucleic acid via weak non-covalent interactions. There are a number of fluorescent probes that change their fluorescent properties (i.e. their quantum yield and/or spectral characteristics) upon association with nucleic acid. Such probes are frequently used to detect, visualize and follow processes involving nucleic acid and its conformational changes. In order to obtain reliable data regarding macromolecule or aggregate topology a detailed knowledge of probe-nucleic acid interactions on the molecular level is needed. In this paper we show that the association of propidium iodide with DNA alters its conformation and that it selectively labels plasmid fragments and/or its subpopulations in a concentration-dependent meaner. Another dye, PicoGreen, exhibits better properties. It labels nucleic acid uniformly and without any concentration-dependent artifacts.
Kral
Teresa
T
Department of Physics and Biophysics, Agricultural University, Norwida 25, 50-375 Wrocław, Poland. tek@ozi.ar.wroc.pl
Widerak
Katarzyna
K
Langner
Marek
M
Hof
Martin
M
D004247:DNA / Q000032:analysis*; D005456:Fluorescent Dyes / Q000737:chemistry*; D009930:Organic Chemicals; D011419:Propidium / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes; D009930:Organic Chemicals; C099476:PicoGreen; D011419:Propidium; D004247:DNA