Molecular size determinations of DNA restriction fragments and polymerase chain reaction products using capillary gel electrophoresis.
Commercially available capillary electrophoresis (CE) systems offer advantages over traditional slab gel methodologies. The capillary format allows the use of higher voltages (225 V/cm), which results in faster migration, higher resolution and greater efficiency without excessive heating. The ability to automate the system increases the unattended sample analysis throughput. For this study, the CE system was configured with a muPAGE 3% T, 3% C polyacrylamide gel capillary with an effective length of 40 cm and microPAGE Tris-borate urea buffer system. The analysis of DNA restriction fragments and polymerase chain reaction products, with an internal standard of Boehringer Mannheim DNA marker XI were completed in less than 70 min. All samples were analyzed at 260 nm. The data establishes automated capillary gel electrophoresis as a high-resolution, reproducible method for analysis of samples under 1000 base pairs.
Marino
M A
MA
Developmental Laboratory, Armed Forces Institute of Pathology, Washington, DC 20306-6000.
Turni
L A
LA
Del Rio
S A
SA
Williams
P E
PE
D001482:Base Composition; D002021:Buffers; D002197:Capillary Action; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D013025:Glycine max / Q000235:genetics
D002021:Buffers; D004247:DNA; D004262:DNA Restriction Enzymes