GM003  6x DNA Loading Buffer

Description: a pre-mixed loading buffer with a tracking dye for agarose and non-denaturing poylacrylamide gel electrophoresis

Final 1x Gel Loading Dye:

2.5 % Ficoll 400

11 mM EDTA [b]

3.3 mM Tris-HCl

0.017 % SDS [a]

0.15 % Orange G [c]

pH 8.0 @ 25°C

Prepare 6x Gel Loading Dye (50 mL):

7.5 g              Ficoll 400

6.6 mL            0.5 M EDTA, pH 8.0

0.5 mL            2 M Tris-HCl, pH 8.0

0.5 mL            10% SDS, pH 8.0

100 mg           Orange G

pH 8.0 @ 25°C

             

Notes: 

[a] This solution contains SDS, which often results in sharper bands, as some restriction enzymes are known to remain bound to their DNA substrates following cleavage.

[b] EDTA is also included to chelate magnesium (up to 10  mM) in enzymatic reactions, thereby stopping the reaction.

[c] Orange G will not appear in gel photographs, and runs ahead of all but the smallest restriction fragments. Orange G migrates at approximately 50 bp on a standard 1% TBE agarose gel.

Reference:  New England Biolabs (NEB) recipe