Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.
Abstract
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.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- August 1988
- DOI:
- 10.1073/pnas.85.15.5610
- Bibcode:
- 1988PNAS...85.5610B