bio-acrylic-resin and Colonic-Neoplasms

cDNA representational difference analysis (cDNA RDA) is a PCR-based subtractive enrichment procedure for the cloning of differentially expressed genes. In this study, we have further developed the procedure to take advantage of solid-phase technology, and to facilitate the use of RDA when starting material is limited. Several parameters of the PCR-based generation of cDNA representations were investigated, and a solid-phase based purification step was introduced to simplify removal of digested adapter-ends and uncleaved fragments. The use of magnetic particles increased the speed of the method, and also eliminated the risk of carry-over contamination between iterative steps of subtraction and PCR amplification. The modified protocol was evaluated in monitoring differences in gene expression in (i) a rat system consisting of livers with and without growth hormone treatment, and in (ii) a human system consisting of normal colon and colon cancer.

Topics: Acrylic Resins; Animals; Biopsy; Cloning, Molecular; Colonic Neoplasms; DNA Primers; DNA, Complementary; DNA, Neoplasm; Electrophoresis, Agar Gel; Growth Hormone; Humans; Liver; Male; Nucleic Acid Hybridization; Polymerase Chain Reaction; Rats; RNA, Messenger; Subtraction Technique; Tissue Extracts