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Automation of dideoxynucleotide DNA sequencing reactions using a robotic workstation.

Authors :
Wilson RK
Yuen AS
Clark SM
Spence C
Arakelian P
Hood LE
Source :
BioTechniques [Biotechniques] 1988 Sep; Vol. 6 (8), pp. 776-7, 781-7.
Publication Year :
1988

Abstract

We report here a system for automation of the dideoxynucleotide DNA sequencing method. The system consists of a Beckman Biomek 1000 robotic workstation which has been modified by the addition of a thin heater/cooler block directly on the instrument table. The heater/cooler block, which is regulated by a user-specified program integrated into the Biomek software, facilitates the use of both single- and double-stranded DNA sequencing procedures. Using this system, we are able to perform 24 sets of dideoxynucleotide DNA sequencing reactions in approximately 45 min. The reactions are performed in 96-well microtiter plates, using a prepared reagent pack which includes primer, enzyme, nucleotide mixes and radioactive label. Using standard gel electrophoresis technology, we are able to resolve over 500 nucleotides per sequencing reaction in about 5 h. Currently, we are able to perform three runs of 24 samples each, with subsequent gel analysis per 8-h period. Excluding autoradiography, this represents a daily data output of 36,000 base pairs.

Details

Language :
English
ISSN :
0736-6205
Volume :
6
Issue :
8
Database :
MEDLINE
Journal :
BioTechniques
Publication Type :
Academic Journal
Accession number :
3273640