Xylene Cyanol FF, an acid triphenylmethane dye, is utilized for histochemical staining of hemoglobin peroxidase and as a tracking dye for DNA sequencing during electrophoresis. Catalyzed by Fe and Al, it accelerates oxidation with hydrogen peroxide and potassium periodate, enabling spectrophotometric determination of Fe and Al in solutions [1] [2].

Xylene Cyanol FF is utilized to trace DNA in various polyacrylamide gel electrophoresis setups. [1] The procedures are as follows: 1. For denaturing gels: (1) Prepare a gel consisting of 10% acrylamide (19:1 acrylamide:bisacrylamide) and 8.3 M urea, running it at 55°C. (2) Prepare the electrophoresis buffer using 89 mM Tris, HCl at pH 8.0, 89 mM boric acid, and 2 mM EDTA (TBE). (3) Prepare the loading buffer with 10 mM NaOH, 1 mM EDTA, and 0.1% Xylene Cyanol FF. (4) Run the gel on an IBI model STS 45 apparatus at 70 W (50 V/cm, constant power) or on a Hoefer SE 600 apparatus at 60°C (31 V/cm, constant voltage). (5) Dry the gel on Whatman 3MM paper and expose it to X-ray film for up to 15 hours. 2. For another denaturing gel method: (1) Prepare a gel containing 8% acrylamide (19:1 acrylamide:bisacrylamide). (2) Make a DNA suspension with 40 mM Tris-HCl at pH 8.0, 20 mM acetic acid, 2 mM EDTA, and 12.5 mM magnesium acetate (TAEMg). (3) Boil the DNA suspension and cool slowly to 16°C. (4) Prepare a dye solution with TAEMg, 50% glycerol, 0.02% bromophenol blue, and 0.02% Xylene Cyanol FF to adjust the sample to a final volume of 10 µL. (5) Run the gel on the Hoefer SE-600 apparatus at 11 V/cm and 16°C, and expose it to X-ray film for up to 15 hours or stain with Stainsall dye.