In:
Applied Spectroscopy, SAGE Publications, Vol. 66, No. 2 ( 2012-02), p. 170-174
Abstract:
The fluorescent small molecule 2-amino-7-methyl-1,8-naphthyridine (AMND) can selectively bind to a cytosine (C) at a C–C mismatch in double-stranded DNA (dsDNA). The interactions between AMND and C–C mismatch-containing dsDNA were investigated by measuring ultraviolet (UV) absorption as a function of temperature to obtain melting curves as well as circular dichroism and fluorescence spectra. Results show that AMND strongly stabilizes C–C mismatch-containing dsDNA, whereas fully matched duplexes are not stabilized under the same conditions. The fluorescence of AMND was efficiently quenched when it was bound to a C–C mismatch in dsDNA. Binding constants ( K 11 ), obtained by fluorescence titration, were 1.2 × 10 5 M −1 . Although sensing functions depend on the sequences flanking the mismatch site, the change in AMND fluorescence intensity can be utilized to detect the C–C mismatch-containing dsDNA. Accordingly, discrimination of the C/G mutation in the model sequence ( PGR gene rs1255998) was achieved by visualizing fluorescence of AMND. A probe DNA molecule was designed to contain a C opposite the C/G base in the target DNA, and this probe was used to hybridize the target DNA. The fluorescence of AMND was “on” for a C–G match, while the fluorescence was “off” for a C–C mismatch. This assay is simple and does not require DNA labeling.
Type of Medium:
Online Resource
ISSN:
0003-7028
,
1943-3530
Language:
English
Publisher:
SAGE Publications
Publication Date:
2012
detail.hit.zdb_id:
1474251-2
SSG:
11
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