Xylene Cyanol FF

Guide (The following is our recommended experimental protocol. This protocol is for reference only and should be modified according to your specific needs).
Xylene Cyanol FF tracks DNA on different polyacrylamide gels. The operation is as follows^[1]:
1. Used for denaturing gel.
(1) Prepare gel: Contain 10% acrylamide (19:1, acrylamide:bisacrylamide), and 8.3 M urea. The operating condition is 55°C.
(2) Prepare electrophoresis buffer: 89 mM Tris, HCl, pH 8.0, 89 mM boric acid, 2 mM EDTA (TBE).
(3) Prepare loading buffer: 10 mM NaOH, 1 mM EDTA, and 0.1% Xylene Cyanol FF.
(4) Gel is run at 70 W (50 V/cm, constant power) on an electrophoresis unit, or 60 °C (31 V/cm, constant voltage).
(5) Dry the gel on Whatman 3MM paper and expose to X-ray film. Exposure time up to 15 hours.
2. Used for denaturing gel.
(1) Prepare the gel: Contain 8% acrylamide (19:1, acrylamide:bisacrylamide).
(2) Prepare DNA suspension: containing 40 mM TrisβHCl, 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 staining solution: containing TAEMg, 50% glycerol, 0.02% bromophenol blue and 0.02% Xylene Cyanol FF. Bring up the sample to a final volume of 10 μL with this staining solution.
(5) The gel is run on a Hoefer SE-600 gel electrophoresis unit at 11 V/cm at 16 °C and exposed to X-ray film for up to 15 hours or stained with Stainsall (HY-D0987).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.