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Nucleic Acids Research, Vol 27, Issue 4 1079-1083, Copyright © 1999 by Oxford University Press

ARTICLES

cDNA fingerprinting of osteoprogenitor cells to isolate differentiation stage-specific genes

GA Candeliere, Y Rao, A Floh, SD Sandler and JE Aubin
Department of Anatomy and Cell Biology, Room 6230 Medical Sciences Building, University of Toronto,1 King's College Circle, Toronto, Ontario M5S 1A8, Canada. tony.candeliere@utoronto.ca

A cDNA fingerprinting strategy was developed to identify genes based on their differential expression pattern during osteoblast development. Preliminary biological and molecular staging of cDNA pools prepared by global amplification PCR allowed discrim-inating choices to be made in selection of expressed sequence tags (ESTs) to be isolated. Sequencing of selected ESTs confirmed that both known and novel genes can be isolated from any developmental stage of interest, e.g. from primitive progenitors, intermediate precursors or mature osteoblasts. EST expression provides insight into possible interrelated physiological functions and putative interacting molecules during differentiation. This method offers a functional genomics approach to isolate differentiation stage-specific genes in samples as small as a single cell.
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