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T7-Based Linear Amplification of Low Concentration mRNA Samples using Beads and Microfluidics for Global Gene Expression Measurements

Published

Author(s)

Jason G. Kralj, Audrey Player, Hope Sedrick, Matt S. Munson, David Petersen, Samuel P. Forry, Paul Meltzer, Ernest Kawasaki, Laurie E. Locascio

Abstract

We show single-cell level linear amplification of gene expression libraries immobilized on a microfluidic packed bed. This is a critical step in the Eberwine process, which is used to amplify messenger RNA (mRNA) for gene expression profiling. Microarray assays comparing microfluidic and benchtop reactions typically showed equivalent levels of gene detection from approximately 5000-9000 transcripts (51% to 97% global gene expression), with concordance between methods and replicates above 90 %. Real-time PCR was used to validate the microarray findings and to estimate the amplification factor at approximately 600.
Citation
Lab on A Chip
Volume
9
Issue
7

Citation

Kralj, J. , Player, A. , Sedrick, H. , Munson, M. , Petersen, D. , Forry, S. , Meltzer, P. , Kawasaki, E. and Locascio, L. (2009), T7-Based Linear Amplification of Low Concentration mRNA Samples using Beads and Microfluidics for Global Gene Expression Measurements, Lab on A Chip, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=832379 (Accessed April 21, 2024)
Created April 7, 2009, Updated February 19, 2017