H. Nakashima et al., 2-DIMENSIONAL ELECTROPHORESIS SOUTHERN TRANSFER METHOD FOR DETECTING HUMAN GENOME VARIABILITY USING A LINE-1 SEQUENCE PROBE, Analytical biochemistry, 227(2), 1995, pp. 319-327
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.