Fluorescence labeling methods for microchannel plate capillary electrophoresis DNA sizing.
DNA analysis on microchannel plate (MCP) capillary electrophoresis platforms will be advanced by the development of new methods for the fluorescence labeling of analytes and standard sizing ladders. Here we evaluate end-labeling of commercially available DNA ladders and polymerase chain reaction (PCR) amplicons with terminal deoxynucleotidyl transferase (TdT) as a method for fluorescent double-stranded DNA sizing analysis. A PCR-based procedure for the facile construction of custom energy-transfer-labeled DNA sizing ladders is also presented. High-resolution sizing of single-stranded DNA fragments is demonstrated with this energy-transfer-labeled ladder. These DNA labeling procedures will be useful for double-and single-stranded DNA analyses on microdevices and other electrophoretic platforms.
Medintz
Igor
I
Center for Biomolecular Science and Engineering, Naval Research Lab, Washington, DC 20375, USA.
Wong
Wendy W
WW
Mathies
Richard A
RA
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D017931:DNA Primers; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes; D008832:Microchemistry; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008970:Molecular Weight
D017931:DNA Primers; D005456:Fluorescent Dyes; D004247:DNA