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BioTechniques
Article . 1996 . Peer-reviewed
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BioTechniques
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BioTechniques
Article . 1997
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BioTechniques
Article . 1996
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High-Throughput Asymmetric-PCR SSCP Analysis Using Well-Controlled Temperature Conditions

Authors: Masaharu Kiyama; Takeshi Fujita;

High-Throughput Asymmetric-PCR SSCP Analysis Using Well-Controlled Temperature Conditions

Abstract

Single-stranded conformational polymorphism (SSCP) analysis is useful for mutation detection and polymorphism identification. To realize high-throughput SSCP analysis using a high-intensity electric field, the gel temperature must be well-controlled. We have developed an electrophoretic system having a pair of glass plates with an efficient temperature-controlling apparatus on each side of the gel. The gel temperature is controlled by estimating the temperature difference between the gel and the glass plates, based on a heat-transfer model. Distinctive patterns of asymmetric-PCR SSCP gel electrophoresis were obtained with this system. The patterns showed high reproducibility for an electric field ranging from 40 V/cm to 160 V/cm, as long as the same gel temperature was used. Electric-field strength had little effect on the SSCP patterns. Use of the highest electric field reduced the run time to about 100 min.

Related Organizations
Keywords

QH301-705.5, HLA-DQ Antigens, Temperature, Humans, Biology (General), Polymerase Chain Reaction, Polymorphism, Single-Stranded Conformational

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    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Average
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
6
Average
Average
Average
gold