Capillary electrophoresis with laser-induced fluorescence detection of PCR fragments using thiazole orange

Structure–Fluorescence Contrast Relationship in Cyanine DNA Intercalators: Toward Rational Dye Design

“…[6] These attractive characteristics enabled DNA to be detected for the first time at a sensitivity comparable to that of radioactive probes, but without the danger inherent in radioactivity. [7] Although some sequence specificity that might lead to inhomogeneous staining has been demonstrated for both TOTO and, to a lesser extent, YOYO, [8][9][10] these cyanines and some derivatives have been used as general DNA stains in numerous DNA detection and quantitation assays, [11] such as the polymerase chain reaction, [12,13] DNA staining and fragment sizing, [14][15][16][17][18] DNA damage detection, [19,20] flow cytometry, [16,21,22] evaluation of biological activity, [23,24] DNA imaging [25][26][27][28] and DNA photocleavage. [29][30][31] YO and TO have also been covalently linked to oligonucleotides and inserted into peptide nucleic acids constructs, which become fluorescent upon hybridisation of the light-up probe to a specific complementary strand.…”

Advances in capillary electrophoresis

“…[194]. The bound dye emisses with a much higher intensity compared to the unbound molecule and a red shift in the emission wavelengths can be observed (520 nm when bound, 580 nm when unbound) The experience obtained with this dye is quite plausible [195].…”

Separation of polymerase chain reaction amplified bird genes by capillary electrophoresis

“…Simple (monomeric) intercalators such as EB and TO, which have reduced steric constraints, could potentially intercalate with a ratio of 1/1 (dye/bp). However, results from previous studies indicated a saturation maximum of one intercalator molecule per two base pairs [29,30]. The intercalating dye is expected to serve a dual role in the separation process.…”