MULTIPLE SEPARATIONS OF DNA SEQUENCING FRAGMENTS WITH A NON-CROSS-LINKED POLYACRYLAMIDE-FILLED CAPILLARY - CAPILLARY ELECTROPHORESIS AT 300 V/CM

摘要

The use of non-cross-linked polyacrylamide for multiple DNA sequencing runs on the same capillary is demonstrated. To minimize template loading, cycle sequencing was used to prepare fluorescently labeled samples. To minimize ion depletion, the current was reversed for several minutes between runs. To achieve stable operation, non-cross-linked polyacrylamide was aged for several days before use. This procedure allowed the successful generation of at least nineteen sequencing runs from the same capillary without replacement of the polyacrylamide and without trimming of the capillary tip. These separations were performed at an electric field of 300 V/cm.