A novel strategy for identifying differential gene expression: An improved method of differential display analysis

We propose a novel alternative approach, an advanced method for recently developed strategies, for identifying differentially expressed genes, Firstly, double-stranded cDNAs were digested using Sau3AI and the 3'-end restriction fragments of the cDNA were ligated to a double-stranded adapter. Next, the restriction fragments were directly amplified using several combinations of adapter-specific primers and FITC-labeled oligo dT primers. The selected cDNA fragments were displayed on a polyacrylamide gel. Neither nested PCR nor purification of 3'-end fragments are necessary. We examined the validity of this approach by evaluating gene expression changes during granulocytic differentiation of HL-60 cells, This method can theoretically detect almost all gene expression changes more rapidly and through simpler manipulations than by any other approach. (C) 1999 Academic Press.