Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.
A method for sequencing DNA by using a difluoresceinated primer and laser excitation is described. Dideoxy protocols have been determined that provide sequences for 600 bases starting with base 1 with less than 1% error in a single load. Electrophoresis is at 20 W and the bands are detected 24 cm from the bottom of the loading well with a scanning fluorescence detector. Bands are imaged on a TV screen in two dimensions. The sequences can be read from the TV screen manually or semiautomatically by using a simple software program. The system allows more bases to be read with a lower error rate than any other reported automated sequencing method.
Brumbaugh
J A
JA
School of Biological Sciences, University of Nebraska, Lincoln 68588-0118.
Middendorf
L R
LR
Grone
D L
DL
Ruth
J L
JL
D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA / Q000235:genetics*; D003857:Deoxyuridine / Q000031:analogs & derivatives; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D009841:Oligonucleotides; D012984:Software
D005456:Fluorescent Dyes; D009841:Oligonucleotides; D004247:DNA; D003857:Deoxyuridine