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- The complementary pairs of single strands were heated in a borate buffer (pH 8.3) for 10 min at 85–90° C, and the mixture was allowed to cool overnight. The duplexes were purified by HPLC on a DIONEX NucleopacTM PA-100 column, then by HPLC on a reverse phase Zorbax SB-C8 column, followed by passage through Sephadex G-25
- Measurements were made at a DNA cone. of 1 mM in a 90 mM Tris-borate buffer (pH 8.3), with 100 mM NaCl and 0.1 mM EDTA (chosen for FRET conditions to maximize fluorescein quantum yield). These studies were repeated in phosphate buffer at pH 7.4 with no change in the results
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