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PMID: 12703299 Published · ppublish English Evaluation Study Research Support, Non-U.S. Gov't Technical Report

Single-stranded conformational polymorphism analysis using automated capillary array electrophoresis apparatuses.

BioTechniques ·Vol. 34 ·No. 4 ·2003-04-00 ·Pages 746-50

Baba S, Kukita Y, Higasa K, Tahira T, Hayashi K

Abstract

We describe a new environment of a single-stranded conformational polymorphism (SSCP) analysis using automated capillary array sequencers (e.g., ABI Prism 3100 and 3700). In this environment, electrophoretic conditions, settings for instrument management, and software for data analysis are adjusted for SSCP analysis. Highly reproducible results are obtained with this new system, and fragments with mutations and/or polymorphisms in different capillaries or different runs can be reliably detected. The relative peak heights between alleles are quantitative and reproducible between runs, and so allele frequencies of single nucleotide polymorphisms can be accurately estimated by a pooled DNA strategy. The method allows unattended, low-cost, and quantitative SSCP analysis using instruments that are widely accessible.

MeSH Terms
Alleles DNA/chemistry,genetics DNA Mutational Analysis/methods Electrophoresis, Capillary/methods Gene Expression Profiling Gene Frequency/genetics Polymerase Chain Reaction/methods Polymorphism, Single-Stranded Conformational Reproducibility of Results Sensitivity and Specificity Sequence Alignment/methods
Chemicals
DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Baba S
Kyushu University, Fukuoka, Japan.
Kukita Y
Higasa K
Tahira T
Hayashi K
Article Info
Journal
BioTechniques
Abbr.
Biotechniques
ISSN
0736-6205
Published
2003-04-00
Pages
746-50
Language
English
Region
England
NLM ID
8306785
Subset
IM
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