The Bacteriophage T4 DNA Ligase Assay Kit Plus (Catalog number T4L100KE) includes 500 ul of 10 x buffer, 44 ul of 100 x DNA substrate, 44 ul of 100 x ATP (2 mM), 21 ml of Reagent T, 220 ul of 10 x dye  and 44 ul of 100 x T4 DNA ligase (5000 U/ml) for 100 assays.