The 5’-Dichloro-dimethoxy-fluorescein dye (JOE) is a traditional fluorophore highly useful for automated DNA sequencing. For Applied Biosystems DNA PRISM sequencers, the dye sets FAM/JOE/TAMRA/ROX and FAM/JOE/NED/ROX can be employed. JOE’s emission falls between the other dyes used in automated sequencing, allowing it to be resolved from FAM and TAMRA. This allows multiplex detection without causing significant signal bleed-over into other channels. It exhibits low sensitivity to pH as the fluorescein dye is less prone to quenching by protonation, accounting for the decrease in signal of mere 30% at pH 5. Additional tributes are high fluorescent quantum yield (number of photons emitted per photon absorbed) of 0.581, high extinction coefficient (5,000 L/mol.cm), and stability under deprotection using ammonium hydroxide.
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-20°C To -70°C Oligonucleotides are stable in solution at 4°C for up to 2 weeks. Properly reconstituted material stored at -20°C should be stable for at least 6 months. Dried DNA (when kept at -20°C) in a nuclease-free environment should be stable for years.
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