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 large-scale analysis, is based on solid-phase DNA sequencing of amplified genomic DNA and does not involve preparative electrophoresis, precipitations, extractions or centrifugations. Here we show that basal cell carcinoma tissue isolated from human skin can be analyzed in a semiautomated fashion using a robotic workstation and a laser fluorescent electrophoresis unit. The results of analyses of patient material displaying various immunohistochemical staining patterns using a p53-specific antibody are presented.