Loading dyes¶

TriTrack¶

Source |150 - Contents: - 10 mM Tris-HCl (pH 7.6) - 0.03 % bromophenol blue - 0.03 % xylene cyanol FF - 0.15 % orange G - 60 % glycerol - 60 mM EDTA. - Uses: - Agarose gel - Native PAGE

Gel Loading Buffer II¶

Source - 1-2X solution - Contents: - 95% Formamide - 18 mM EDTA - 0.025% each of SDS, Xylene Cyanol, and Bromophenol Blue - Uses: - Polyacrylamide urea gel (denaturing) - Non-denaturing agarose gel - Appearance: Dark blue

Protocol: 1. Mix sample with an equal volume of Gel Loading Buffer II. Vortex briefly. 2. Centrifuge briefly to bring contents of tubes to the bottom. 3. For denaturing PAGE: Heat to 95°C for 5 min to denature any secondary structure. 4. Load directly (while still hot) on the gel.

RNA Loading Dye¶

Source - 2X - Contents: - 47.5% Formamide - 0.01% SDS - 0.01% bromophenol blue - 0.005% xylene cyanol - 0.5 mM EDTA - Uses: - Polyacrylamide urea gel (denaturing)

Protocol: 1. Add sample to an equal volume of RNA Loading Dye, (2X). Mix well. 2. Heat at 65–70°C for 5–10 minutes to denature RNA. 3. While heating the samples, setup the gel box and flush urea out of the wells with running buffer using a large tip. 4. Load samples.