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Although agarose gels can easily be prepared in an open-faced format to gain
the conveniences of horizontal electrophoresis, acrylamide does not polymerize
in the presence of air and the usual configurations
for gel preparation lead to electrophoresis
in the vertical dimension. We describe here a very simple device and method to
prepare and manipulate horizontal polyacrylamide gels (H-PAGE). In addition,
the open-faced horizontal arrangement enables loading arrays of wells. Since
many procedures are undertaken in standard 96-well microtiter plates, we have
also designed a device which preserves the exact configuration of the 8 x 12
array and enables electrophoresis in
tracks following a 71.6 degree diagonal between wells (MADGE, microtiter
array diagonal gel electrophoresis),
using either acrylamide or agarose. This eliminates almost
all of the staff time taken in setup, loading and record keeping and offers high
resolution for genotyping pattern recognition. The nature and size of the gels
allow direct stacking of gels in one tank, so that a tank used typically to analyze 30
- 60 samples can readily be used to analyze 1000 - 2000 samples. The
gels would also enable robotic loading.
Electrophoresis allows
analysis of size and charge, parameters inaccessible to liquid-phase methods:
thus, genotyping size patterns, variable length repeats, and haplotypes
is possible, as well as adaptability to typing of point variations using
protocols which create a difference detectable by electrophoresis
ANALYTICAL BIOCHEMISTRY 222, 389-395 (1994)
