Automated DNA mutation analysis by single-strand conformation polymorphism using capillary electrophoresis with laser-induced fluorescence detection.

M S Liu; S Rampal; D Hsiang; F T Chen

doi:10.1385/mb:15:1:21

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PubMed · 10911619

Automated DNA mutation analysis by single-strand conformation polymorphism using capillary electrophoresis with laser-induced fluorescence detection.

M S Liu·S Rampal·D Hsiang·F T Chen

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Abstract

Automation is essential for rapid genetic-based mutation analysis in clinical laboratory to screen a large number of DNA samples. We propose in this report an automatic process using Beckman Coulter P/ACE capillary electrophoresis (CE) with laser-induced fluorescence (LIF) system to detect a single-point mutation in the codon 12 of human K-ras gene. Polymerase chain reaction (PCR) using a fluorescently labeled reverse primer and a plain forward primer to specifically amplify a selected 50 bp DNA fragment in human K-ras gene. The amplified DNA is placed on the sample tray of the CE system with a pre-programmed step for single-strand conformation polymorphism (SSCP) analysis. Sample injection and denaturation processes are performed online along with separation and real-time data analysis. The concept of automation for rapid DNA mutation analysis using CE-LIF system for SSCP is presented.

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Automation Base Sequence DNA DNA Mutational Analysis Electrophoresis, Capillary Fluorescence Genes, ras Humans Lasers Molecular Sequence Data Point Mutation Polymerase Chain Reaction Polymorphism, Single-Stranded Conformational Sodium Hydroxide Journal Article

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M S Liu, S Rampal, D Hsiang, F T Chen. 2000. Automated DNA mutation analysis by single-strand conformation polymorphism using capillary electrophoresis with laser-induced fluorescence detection.. https://doi.org/10.1385/mb%3A15%3A1%3A21

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