Nucleic Acids Research, Vol 24, Issue 8 1435-1442, Copyright © 1996 by Oxford University Press
K Bernard, N Auphan, S Granjeaud, G Victorero, AM Schmitt-Verhulst, BR Jordan and C Nguyen
The hybridization signature approach, using colony filters and labeled
complex probes, can provide high throughput measurement of gene activity.
We describe here the implementation of this method to follow the expression
levels of 47 genes in resting and activated T cells, as well as in
epithelial cells. Using 4-fold spotting of colonies, imaging plate
detection and various correction and normalization procedures, the
technique is sensitive enough to quantify expression levels for sequences
present at 0.005% abundance in the probe. Comparison with Northern blotting
shows good consistency between the two methods. Upon activation of a T cell
clone by an anti-CD3 antibody variations ranging from 2- to 20-fold are
measured, some of which had not been reported previously. This 'multiplex
messenger assay' method, performed using available commercial apparatus,
can be used in many cases where simultaneous assessment of mRNA levels for
many genes is of interest.
ARTICLES
Multiplex messenger assay: simultaneous, quantitative measurement of expression of many genes in the context of T cell activation
Genome Structure and Immune Functions laboratory, Marseille, France.
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