The interactions between the fluorescent dye thiazole orange and DNA

Nygren, Jan; Svanvik, Nicke; Kubista, Mikael

doi:10.1002/(sici)1097-0282(199807)46:1<39::aid-bip4>3.0.co;2-z

Cited by 419 publications

(163 citation statements)

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“…A similar red-shift is observed for monomeric TO bound to DNA ( max TO-DNA ϭ 507-511) (16). Some of the free probe spectra also exhibit shoulders around 485 nm that are not observed for free TO (Fig.…”

Section: Fluorescence Of Free Light-up Probessupporting

confidence: 71%

“…They also have the smallest shift in absorption ( max 25°C ϭ 506 -507 nm) relative to free TO ( max 25°C ϭ 501 nm). This suggests that TO in these probes is mainly free in solution and does not interact significantly with the PNA, which is consistent with the generally very low affinity of TO for polypyrimidines (16). The variation in free probe fluorescence of probes containing all four nucleobases is considerably larger (⌽ F 25°C ϭ 0.0024 -0.083).…”

Section: Figsupporting

confidence: 69%

“…Some of the free probe spectra also exhibit shoulders around 485 nm that are not observed for free TO (Fig. 2), but appear when TO binds to DNA (16). Further, the free probes exhibit higher fluorescence than free TO ( Table 2).…”

Section: Fluorescence Of Free Light-up Probesmentioning

confidence: 90%

“…Figure 2 shows absorption spectra of LUP2, LUP3, and of free TO at 25°C. Free TO absorbs mainly in the VIS region having absorption maximum at 501 nm and a molar absorptivity of 63,000 M Ϫ1 cm Ϫ1 (16). Both TO ( max LUP2 ϭ 512, max LUP3 ϭ 506 nm) and PNA ( max LUP2 ϭ 260, max LUP3 ϭ 270 nm), whose absorption depends on the sequence, contribute to the probe absorption.…”

Section: Tablementioning

confidence: 99%

“…At 25°C both these probes are more fluorescent than free TO, having fluorescence quantum yields (⌽ F ) of 0.0089 (LUP2) and 0.0015 (LUP3), while free TO has 0.0002 (16). The spectroscopic properties of the light-up probes are summarized in Table 2.…”

Section: Tablementioning

confidence: 99%

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Light-Up Probes: Thiazole Orange-Conjugated Peptide Nucleic Acid for Detection of Target Nucleic Acid in Homogeneous Solution

Svanvik

Westman

Wang³

et al. 2000

Analytical Biochemistry

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We have constructed light-up probes for nucleic acid detection. The light-up probe is a peptide nucleic acid (PNA) oligonucleotide to which the asymmetric cyanine dye thiazole orange (TO) is tethered. It combines the excellent hybridization properties of PNA and the large fluorescence enhancement of TO upon binding to DNA. When the PNA hybridizes to target DNA, the dye binds and becomes fluorescent. Free probes have low fluorescence, which may increase almost 50-fold upon hybridization to complementary nucleic acid. This makes the light-up probes particularly suitable for homogeneous hybridization assays, where separation of the bound and free probe is not necessary. We find that the fluorescence enhancement upon hybridization varies among different probes, which is mainly due to variations in free probe fluorescence. For eight probes studied the fluorescence quantum yield at 25°C in the unbound state ranged from 0.0015 to 0.08 and seemed to depend mainly on the PNA sequence. The binding of the light-up probes to target DNA is highly sequence specific and a single mismatch in a 10-mer target sequence was readily identified.

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Section: Fluorescence Of Free Light-up Probessupporting

confidence: 71%

Section: Figsupporting

confidence: 69%

Section: Fluorescence Of Free Light-up Probesmentioning

confidence: 90%

Section: Tablementioning

confidence: 99%

Section: Tablementioning

confidence: 99%

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