UNIT 25B.8 Gene Expression Analysis of a Single or Few Cells
Published Online: 1 FEB 2003
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Klein, C. A., Zohlnhöfer, D., Petat-Dutter, K. and Wendler, N. 2003. Gene Expression Analysis of a Single or Few Cells. Current Protocols in Molecular Biology. 61:B:25B.8:25B.8.1–25B.8.18.
- Published Online: 1 FEB 2003
- Published Print: JAN 2003
The need to analyze rare cells is based on the nature of tissue differentiation and regeneration, the initiation and propagation of disease processes in multicellular organisms, and the functional diversity of individual cells. Gene transcription is the most important regulatory mechanism by which a phenotype and functional state of a cell is determined. Therefore, procedures for the qualitative and quantitative assessment of mRNA abundance are important. This unit presents a protocol for semi-quantitative analysis of gene expression of a single cell and quantitative representation of expressed genes from >10 to 30 cells. A basic protocol for array hybridization on nylon filters is provided because such filters are available in every laboratory. Tissue samples contain many different cell types in variable amounts, so their analysis may require microdissection; a protocol for obtaining cryosections is given. Finally, a simple procedure to prepare the data for statistical analysis is also provided.