Reference dye for fluorescence signal normalization
For in vitro use only!
Shipping: shipped on blue ice
Storage Conditions: store at -20 °C
Shelf Life: 12 months
Form: liquid (Supplied in 20 mM Tris-HCl pH 8.4, 0.1 mM EDTA and 0.01 % Tween-20)
Concentration: 10 μM (100x conc.)
Spectroscopic Properties: λexc 485 nm; λem 514 nm; ε 93.0 L mmol-1 cm-1 (pH 9.0); quantum yield 0.93 (pH 9.5)
Fluorescein Reference Dye is optimized for block-based real-time PCR instruments that are compatible with the evaluation of a Fluorescein reference signal. The dye does not take part in the qPCR reaction but allows to normalize for non-PCR related signal variation and provides a baseline in multiplex reactions. The dye is supplied as 100x concentration.
Preparation of the qPCR master mix:
The preparation of a master mix is crucial in quantitative PCR reactions to reduce pipetting errors. Vortex the Fluorescein Reference Dye thoroughly prior to addition to the master mix. Add 0.5 μl Fluorescein Reference Dye for each 50 μl of master mix.
A fluorescein concentration of 100 mM in the final assay is recommended.