High resolution-separation of DNA bands by electrophoresis with a long gel in a fluorescence-detection DNA sequencer.
A high-resolution separation of DNA bands is achieved by electrophoresis with a long gel in DNA base sequencing using fluorescence detection. We separate 760 and 761 base DNA fragments using the 93 cm migration electrophoresis optimized for the separation of DNA bands. A T7 DNA polymerase and an Mn++ buffer are used in sequencing reactions to obtain fluorescence peaks of uniform strength, and the peak areas in the spectrum are used for recognizing the peak number in a cluster of successive peaks. This method is successfully applied to the DNA fragment spectrum obtained by 93 cm migration electrophoresis, which results in a single-band differentiation of bands of 1040 base DNA.
Nishikawa
T
T
Central Research Laboratory, Hitachi Ltd., Tokyo, Japan.
Kambara
H
H
D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D013050:Spectrometry, Fluorescence
D004247:DNA