SEQUENCE-BASED ANALYSIS OF THE HUMAN P53 GENE BASED ON MICRODISSECTION OF TUMOR-BIOPSY SAMPLES

A procedure for direct sequencing of the human p53 gene based on micro -dissection of frozen tumor tissue stained with methylene blue without prior fixation is described. The approach, which is suitable for larg e-scale analysis, is based on solid-phase DNA sequencing of amplified genomic DNA and does not involve preparative electrophoresis, precipit ations, extractions or centrifugations. Here we show that basal cell c arcinoma tissue isolated from human skin can be analyzed in a semiauto mated fashion using a robotic workstation and a laser fluorescent elec trophoresis unit. The results of analyses of patient material displayi ng various immunohistochemical staining patterns using a p53-specific antibody are presented.