Enzyme-linked fluorescent detection for automated multiplex DNA sequencing.
Genomics
; 20(1): 68-74, 1994 Mar 01.
Article
in En
| MEDLINE
| ID: mdl-8020958
ABSTRACT
Initiatives to sequence DNA on a large scale have created a need for increased throughput and decreased costs. One scheme for increasing throughput, multiplex sequencing, involves the processing of a mixture of sequencing templates followed by sequential hybridization to reveal the individual sequence ladders on a membrane. Because multiplex sequencing has not been fully automated, and has not seemed automatable, few sequencing efforts have attempted to exploit it. We describe here a scheme for the automation of multiplex sequencing. Probe hybridized to target DNA is detected via spatially localized enzyme-linked fluorescence. Light output is high enough that imaging is possible with simple instrumentation. Direct imaging within an automated hybridization apparatus is made feasible so that the entire process will be automatic once a multiplex membrane is produced. The technique has the potential to increase severalfold the throughput of automated sequencing instruments required for sequencing the human genome.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Sequence Analysis, DNA
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Journal:
Genomics
Journal subject:
GENETICA
Year:
1994
Document type:
Article