2-DIMENSIONAL ELECTROPHORESIS SOUTHERN TRANSFER METHOD FOR DETECTING HUMAN GENOME VARIABILITY USING A LINE-1 SEQUENCE PROBE

A ''high-resolution, two-dimensional Southern transfer'' method has be en developed and was used to examine the distribution of a class of in terspersed repeated sequences in human genomes. This method consists o f two separate restriction enzyme digestions, including an in situ dig estion, and two dimensional electrophoresis using a large-sized agaros e gel. The first 163-base-pair region of the human LINE-1 full-length sequence was used to probe human genomic DNA from placental tissue sam ples. About 900 LlNE-1 signals were resolved from each DNA sample with in a 2-D plane, The bulk of the fragments were between 0.5 to 23 kilob ases in length, At a minimum 15 variant signals were detected between DNA samples from male and female individuals and at a minimum 16 varia nt signals were detected between two different female samples. This ap proach can potentially be used to perform high-resolution human genome fingerprinting analyses. (C) 1995 Academic Press, Inc.