Agarose and Polyacrylamide Gels

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Agarose and Polyacrylamide Gels

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Adapted from Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). In: Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring ...

Agarose and Polyacrylamide Gels Dye Migration: Polyacrylamide Denaturing Gels.

TECHNICAL REFERENCE

Dyes will migrate to the same point as double-stranded DNA of the indicated size in a denaturing polyacrylamide gel. Gel % 5.0 6.0 8.0 10.0 20.0

Bromophenol Blue 35bp 26bp 19bp 12bp 8bp

Xylene Cyanol 140bp 106bp 75bp 55bp 28bp

Adapted from Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) In: Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

Dye Migration: Polyacrylamide Nondenaturing Gels. Dyes will migrate to the same point as double-stranded DNA of the indicated size in a nondenaturing polyacrylamide gel. Gel % 3.5 5.0 8.0 12.0 15.0 20.0

Bromophenol Blue 100bp 65bp 45bp 20bp 15bp 12bp

Xylene Cyanol 460bp 260bp 160bp 70bp 60bp 45bp

Adapted from Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) In: Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

Dye Migration in 0.5–1.4% Agarose Gels. Dyes will migrate to the same point as double-stranded DNA of the indicated size in an agarose gel. Sizes are approximate. Gel GoTaq® blue dye Xylene cyanol FF Bromophenol Blue Orange G GoTaq® yellow dye

Size 4kb 4kb 300bp 50bp 10bp

Some information adapted from Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) In: Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

Gel Percentages: Resolution of Linear DNA on Agarose Gels. Optimum Resolution for Linear DNA (Size of fragments in base pairs) 1,000–30,000 800–12,000 500–10,000 400–7,000 200–3,000 50–2,000

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Gel Percentages: Resolution of Proteins on Polyacrylamide Gels.

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Recommended % Agarose 0.5 0.7 1.0 1.2 1.5 2.0

Recommended % Acrylamide 8 10 12

Protein Size Range 40–200kDa 21–100kDa 10–40kDa

Dalton (Da) is an alternate name for the atomic mass unit, and kilodalton (kDa) is 1,000 daltons. Thus a peptide with a mass of 64kDa has a molecular weight of 64,000 grams per mole.