doi: 10.1073/pnas.85.15.5610.
Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores
Affiliations
- PMID: 3399506
- PMCID: PMC281809
- DOI: 10.1073/pnas.85.15.5610
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Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores
Proc Natl Acad Sci U S A.
1988 Aug.
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.
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