Simple miniaturized gel system for DNA sequence analysis.
A simple miniaturized gel system suitable for DNA sequencing is described. Small ultrathin polyacrylamide gels are cast, eight or more at a time, using standard microscope slides. Gels, ready to use, can be stored for approximately 2 weeks. Gels are run horizontally in a standard mini-agarose gel apparatus. Typical run times are 6-8 min. A novel sample loading system permits volumes of standard sequencing reactions as small as 0.1 microl to be analyzed. Sequencing ladders were visualized using 35S-labeled DNA by autoradiography and by colorimetric detection. Band resolution compares favorably with that of large gels. The methods introduced here serve as a step toward the miniaturization of DNA sequencing and are amenable to automated sample loading and detection.
Stein
A
A
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA. astein@bilbo.bio.purdue.edu
Hill
S A
SA
Cheng
Z
Z
Bina
M
M
D001345:Autoradiography; D001483:Base Sequence; D003124:Colorimetry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008904:Miniaturization; D017422:Sequence Analysis, DNA / Q000379:methods*; D013462:Sulfur Radioisotopes
D013462:Sulfur Radioisotopes