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PubMed · 10574189

Rapid method for relative gene expression determination in human tissues using automated capillary gel electrophoresis and multicolor detection.

E Odin·Y Wettergren·L Larsson·P A Larsson·B Gustavsson
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Abstract

The aim of this study was to evaluate a direct and automated post-polymerase chain reaction (PCR) detection system to simultaneously determine the relative gene expression levels of nine cancer-related human genes. Total RNA was prepared from flash-frozen biopsies derived from human colorectal tumors or normal mucosa and reverse-transcribed to cDNA which was PCR-amplified using primer pairs corresponding to the studied genes. In each reaction, the forward primer was labeled with a fluorescent dye. The PCR products were pooled and an internal size standard with a uniquely colored fluorescent dye was added. The samples were then subjected to automated capillary gel electrophoresis. Fragment analysis software was used to calculate the relative gene expression using beta-actin as the reference gene. We found that automated capillary gel electrophoresis with multicolor detection is a rapid, accurate and highly reproducible method for separation and quantification of PCR-amplified cDNA.

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AutoanalysisColonColorectal NeoplasmsDNA, ComplementaryDNA, NeoplasmElectrophoresis, CapillaryFluorescent DyesGene ExpressionHumansIntestinal MucosaRectumReproducibility of ResultsReverse Transcriptase Polymerase Chain ReactionSensitivity and SpecificityThymidylate SynthaseJournal ArticleResearch Support, Non-U.S. Gov't

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E Odin, Y Wettergren, L Larsson, P A Larsson, B Gustavsson. 1999-10-29. Rapid method for relative gene expression determination in human tissues using automated capillary gel electrophoresis and multicolor detection.. https://doi.org/10.1016/s0378-4347(99)00333-3

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