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Systematic comparison of gene expression through analysis of cDNA fragments within or near to the protein-coding
1Department of Pathology, Faculty of Medicine and School of Biological Sciences, Liverpool University, Liverpool L69 3BX, UK. yqk@liv.ac.uk
Abstract:
Life is controlled by the timely and ordered expression of genes. Identification of important genes involved in specific physiological and pathological conditions requires efficient methods to analyse differential gene expression. We describe a novel strategy, namely complete comparison of gene expression (CCGE), for a systematic assessment of differentially expressed genes. Using the CCGE method, double-stranded cDNA is digested with two restriction enzymes that cut with different frequencies, the representative cDNA fragments are generated within or near to the protein-coding region. After being flanked by two different types of adapters, and amplified by a nested suppression PCR, the selected cDNA fragments, representing entire cDNA population, can be divided into 256 subsets; amplified and compared in a systematic manner.