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Technical aspects of mRNA quantification in single cells using RT-qPCR

Magazine article
Authors Anders Ståhlberg
Published in International Drug Discovery
Issue April/May
Pages 27-29
Publication year 2010
Published at Institute of Biomedicine, Department of Pathology
Pages 27-29
Language en
Subject categories Cell and Molecular Biology

Abstract

Individual cells are in many aspects unique in their characteristics. Gene expression is highly variable among individual cells, even within a seemingly homogeneous cell population. The desire to uncover fundamental information about cell types, cell heterogeneity and cellular mechanism motivates studying the expression of multiple genes simultaneously in individual cells. Reverse transcription quantitative real-time PCR (RT-qPCR) is the most versatile method that provides sufficiently accurate measurements of mRNA levels in single cells. The low number of transcripts being measured makes good laboratory practice and carefully optimized protocols for single-cell collection, cell lysis, reverse transcription, real-time PCR and data analysis essential for accurate single-cell gene expression profiling. Single-cell gene expression profiling using RT-qPCR has the potential to become common practice in many laboratories and a powerful research/ diagnostic tool for deeper understanding of molecular mechanisms and diseases.

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