Semiconductor-controlled contour-clamped homogeneous electric field apparatus.
The design and construction of a transistor-driven hexagonal contour-clamped homogeneous electric field (CHEF) apparatus is discussed in detail. The addition of computer control of pulsed-field timings and experiment duration gives rise to an efficient electrophoresis tool designed to achieve separation of DNA molecules in different size groupings. In particular, pulse time regimes which lead to the monotonic separation of DNA molecules ranging from 90 kbp to over a megabase pair are demonstrated. Theoretical treatment of electric field clamping with transistor-driven multiple electrodes is supported by measurements and by the actual performance of electrophoretic separation of yeast chromosomes. The large sample capacity of gels run in this apparatus coupled with the modest power requirements necessary to provide a homogeneous electric field offer significant advantages over earlier CHEF designs.
Maule
J
J
MRC Human Genetics Unit, Western General Hospital, Edinburgh, Scotland.
Green
D K
DK
D002021:Buffers; D015825:Chromosomes, Fungal / Q000737:chemistry*; D003201:Computers; D004271:DNA, Fungal / Q000302:isolation & purification*; D004553:Electric Conductivity; D004566:Electrodes; D004574:Electromagnetic Fields; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D002021:Buffers; D004271:DNA, Fungal