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Pulsed-field gel electrophoresis via transverse alternating field electrophoresis gels

Nature Methods volume 3, pages 233–234 (2006)Cite this article

In this form of pulsed-field gel electrophoresis (transverse alternating field electrophoresis, also known as TAFE), electrodes are positioned on opposite sides of a vertical gel1. The DNA moves first toward one electrode and then toward the other, forming a zigzag pattern. The vector of this oscillation is a straight line from the loading well to the base of the gel. Variation in voltage and pulse time allows separation of DNAs ranging in size from 2 kilobases (kb) to >6,000 kb. For the resolution of larger DNA fragments contour-clamped homogeneous electric field gel electrophoresis (CHEF)2 is recommended. This protocol describes the resolution of genomic DNA by TAFE, followed by blotting and hybridization.

1 | Cast a 1% agarose gel in 1× TAFE gel buffer (20 mM Tris-acetate (pH 8.2), 0.5 mM EDTA) without ethidium bromide and allow the gel to set. Run the gel in the same buffer solution as used for preparing the gel.

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References

  1. Gardiner, K. & Patterson, D. Transverse alternating field electrophoresis and applications to mammalian genome mapping. Electrophoresis 10, 296–302 (1989).

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  2. Chu, G., Vollrath D. & Davis, R. Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science 234, 1582–1585 (1986).

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Pulsed-field gel electrophoresis via transverse alternating field electrophoresis gels. Nat Methods 3, 233–234 (2006). https://doi.org/10.1038/nmeth0306-233

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