Fifty EraCodes were validated by use of 50 extenders, all containing the identical 3Ј target-specific sequence. The extenders were individually labeled using the target, standard concentrations of all four naturally occurring deoxynucleotide triphosphates, TiTaq, and diGTP-biotin at 1.5 mol/L. Each EraCode sequence was separately coupled to a specific color-coded fluorescent microsphere type (Luminex Corp). Each microsphere type contains a unique color or signature that can be determined by the Luminex 100 . After coupling, each EraCode bead conjugate was pooled to create an equalized bead master mixture (bead array: 50 different EraCode-labeled beads). The bead array was distributed into 50 separate reaction vessels. To each of the vessels, a specific extension reaction from above was added at room temperature along with streptavidin-phycoerythrin and was read on the Luminex 100 .