Characterization of SYBR gold nucleic acid gel stain: A dye optimized for use with 300-nm ultraviolet transilluminators

被引:250
作者
Tuma, RS [1 ]
Beaudet, MP [1 ]
Jin, XK [1 ]
Jones, LJ [1 ]
Cheung, CY [1 ]
Yue, S [1 ]
Singer, VL [1 ]
机构
[1] Mol Probes Inc, Eugene, OR 97402 USA
关键词
SYBR; nucleic acid; gel stain; fluorescence; UV transillumination;
D O I
10.1006/abio.1998.3067
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The highest sensitivity nucleic acid gel stains developed to date are optimally excited using short-wavelength ultraviolet or visible light, This is a disadvantage for laboratories equipped only with 306- or 312-nm UV transilluminators. We have developed a new unsymmetrical cyanine dye that overcomes this problem. This new dye, SYBR Gold nucleic acid gel stain, has two fluorescence excitation maxima when bound to DNA, one centered at similar to 300 nm and one at similar to 495 nm. We found that when used with 300-nm transillumination and Polaroid black-and-white photography, SYBR Gold stain is more sensitive than ethidium bromide, SYBR Green I stain, and SYBR Green II stain for detecting double-stranded DNA, single-stranded DNA, and RNA. SYBR Gold stain's superior sensitivity is due td the high fluorescence quantum yield of the dye-nucleic acid complexes (similar to 0.7), the dye's large fluorescence enhancement upon binding to nucleic acids (similar to 1000-fold), and its capacity to more fully penetrate gels than do the SYBR Green gel stains. We found that SYBR Gold stain is as sensitive as silver staining for detecting DNA-with a single-step staining procedure. Finally, we found that staining nucleic acids with SYBR Gold stain does not interfere with subsequent molecular biology protocols. (C) 1999 Academic Press.
引用
收藏
页码:278 / 288
页数:11
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