Gel loading buffer is used as a tracking dye during electrophoresis. The dye has a slight negative charge and will migrate the same direction as DNA, allowing the user to monitor the progress of molecules moving through the gel. The rate of migration varies with gel composition. Dilute 1:3 to 1:6 with sample before loading.
Gel Loading Buffer has been used for loading polymerase chain reaction (PCR) amplified intergenic spacer region (ISR) sequence samples, PCR amplified DNA samples of the intestinal mucosa, Trypanosoma sp DNA on agarose gel for electrophoresis.It is suitable for use with agarose or non-denaturing polyacrylamide gel electrophoresis (PAGE), which may be part of Northern and Southern blot hybridization procedures.
Gel loading buffer is especially formulated for non-denaturing polyacrylamide and agarose gel electrophoresis of nucleic acids. The gel loading solution contains a bromophenol blue tracking dye and sucrose to add density and facilitate sample loading. EDTA has been included to inhibit nucleases that require divalent cations and SDS has been added to help dissociate DNA-protein complexes which can interfere with electrophoresis.
Gel loading buffer contains 0.05% bromophenol blue , 40% sucrose, 0.1M EDTA (pH 8.0) and 0.5% SDS.
If crystals form, dissolve them by heating the solution to 65C for 10 minutes.