کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10771897 | 1050845 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Effects of fluorescent dyes, quenchers, and dangling ends on DNA duplex stability
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
Single and dual-labeled fluorescent oligodeoxynucleotides are used in many molecular biology applications. We investigated the effects of commonly used fluorescent dyes and quenchers on the thermodynamic stability of a model probe-target DNA duplex. We demonstrate that those effects can be significant. Fluorescent dyes and quenchers were attached to the probe ends. In certain combinations, these groups stabilized the duplex up to 1.8 kcal/mol and increased Tm up to 4.3 °C. None of the groups tested significantly destabilized the duplex. Rank order of potency was, starting with the most stabilizing group: Iowa Black RQ â¼Â Black Hole 2 > Cy5 â¼Â Cy3 > Black Hole 1 > QSY7 â¼Â Iowa Black FQ > Texas Red â¼Â TAMRA > FAM â¼Â HEX â¼Â Dabcyl > TET. Longer linkers decreased stabilizing effects. Hybridizations to targets with various dangling ends were also studied and were found to have only minor effects on thermodynamic stability. Depending on the dye/quencher combination employed, it can be important to include thermodynamic contributions from fluorophore and quencher when designing oligonucleotide probe assays.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 327, Issue 2, 11 February 2005, Pages 473-484
Journal: Biochemical and Biophysical Research Communications - Volume 327, Issue 2, 11 February 2005, Pages 473-484
نویسندگان
Bernardo G. Moreira, Yong You, Mark A. Behlke, Richard Owczarzy,