SEPARATION OF LARGE DNA FRAGMENTS BY CAPILLARY ELECTROPHORESIS UNDER PULSED-FIELD CONDITIONS

Capillary zone electrophoresis (CZE), using an entangled polyacrylamid e solution, was applied to large DNA samples under pulsed-field condit ions. Highly efficient separations were achieved under biased sinusoid al field and field-inversion pulsing regimes. The separations that wer e obtained with 8.3-48.5 kb lambda DNA standards and 48.5 kb-1 Mb lamb da DNA concatamers (modified with ethidium bromide) clearly demonstrat e a dramatic improvement in the separation time (roughly, 10-50 times) over the conventionally used slab-gel techniques. Moreover, the CZE m ethod appears much more sensitive and amenable to component quantifica tion and method automation.