Direct determination of single-to-double stranded DNA ratio in solution using steady-state fluorescence measurements.
We report a simple and rapid method for quantitation of single-to-double stranded (ss : ds) DNA ratios in solution, using steady-state measurements of fluorescence from two simultaneously excited intercalated dyes; the ratio of fluorescence intensities from PicoGreen (525 nm) and ethidium bromide (610 nm) is directly proportional to the ss : ds DNA ratio.
Beach
Lori
L
Department of Chemistry, University of Ottawa, 10 Marie Curie, Ottawa, Ontario, Canada K1N 6N5.
Schweitzer
Claude
C
Scaiano
J C
JC
D000818:Animals; D002417:Cattle; D004247:DNA / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000032:analysis*; D004996:Ethidium / Q000032:analysis / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D009930:Organic Chemicals; D012996:Solutions / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods
D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C099476:PicoGreen; D012996:Solutions; D004247:DNA; D004996:Ethidium