Molecular Techniques and Methods

Gel Loading Buffers

Copy Right © 2001/ Institute of Molecular Development LLC

  • 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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