Abstract
Determining the chromosomal origin of expressed sequence tags (ESTs) (1,2) lags far behind their identification in single-pass sequencing projects (1–10). Positional cloning of disease genes requires that previously uncharacterized transcripts be mapped to the smallest possible defined region. We have developed an efficient polymerase chain reaction (PCR)-based procedure for the rapid assignment of ESTs to human chromosome regions (11–12; Fig. 1). The critical features of the method are:
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1.
Standard, restricted criteria for primer design;
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2.
Sensitive, automated analysis of fluorescently labeled PCR products,
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3.
Standard PCR conditions; and
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4.
Multiplexed PCR reactions.
Keywords
- Polymerase Chain Reaction Reaction
- Rapid Assignment
- Chromosome Assignment
- Mapping Panel
- Efficient Polymerase Chain Reaction
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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© 1997 Humana Press Inc., Totowa, NJ
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Durkin, A.S., Maglott, D.R., Nierman, W.C. (1997). Mapping Expressed Sequence Tags (ESTs) by Multiplexing PCR Reactions from Hybrid Cell Panels and Detecting Fluorescently Labeled Products. In: Boultwood, J. (eds) Gene Isolation and Mapping Protocols. Methods in Molecular Biology™, vol 68. Humana Press. https://doi.org/10.1385/0-89603-482-8:159
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DOI: https://doi.org/10.1385/0-89603-482-8:159
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