Molecular Techniques and Methods

Gel Loading Buffers

Gel loading buffers increase the density of the sample, ensuring that the DNA drops evenly into the well.

Gel loading buffers add color to the sample.

Gel loading buffers contain dyes that, in an electric field, move toward the anode at predictable rates.

Bromophenol blue migrates through agarose gels run in 0.5 x TBE at approximately the same rate as linear double-stranded DNA 300 bp in length, whereas xylene cyanol FF migrates at approximately the same rate as linear double-stranded DNA 4 kb in length. These relationships are not significantly affected by the concentration of agarose in the gel over the range of 0.5% to 1.4%.

6 x Concentrated Gel Loading Buffer I 0.25% Bromophenol blue 0.25% Xylene cyanol FF 15% Ficoll (Type 400) Distilled H₂O to adjust volume	6 x Concentrated Gel Loading Buffer II 0.25% Bromophenol blue 0.25% Xylene cyanol FF 40% (w/v) Sucrose Distilled H₂O to adjust volume
6 x Concentrated Gel Loading Buffer III 0.25% Bromophenol blue 0.25% Xylene cyanol FF 30% Glycerol Distilled H₂O to adjust volume	6 x Concentrated Gel Loading Buffer IV 0.25% Bromophenol blue 40% (w/v) Sucrose Distilled H₂O to adjust volume
6 x Concentrated Gel Loading Buffer V Alkaline Loading Buffer 300 mM NaOH 6 mm EDTA 18% Ficoll (Type 400) 0.15% Bromocresol green 0.25% Xylene cyanol FF Distilled H₂O to adjust volume Bromocresol green displays a more vivid color than bromophenol blue at alkaline pH.

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