Two-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 been developed and was used to examine the distribution of a class of interspersed repeated sequences in human genomes. This method consists of two separate restriction enzyme digestions, including an in situ digestion, and two-dimensional electrophoresis using a large-sized agarose 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 samples. About 900 LINE-1 signals were resolved from each DNA sample within a 2-D plane. The bulk of the fragments were between 0.5 to 23 kilobases in length. At a minimum 15 variant signals were detected between DNA samples from male and female individuals and at a minimum 16 variant signals were detected between two different female samples. This approach can potentially be used to perform high-resolution human genome fingerprinting analyses.
Nakashima
H
H
Department of Biochemistry, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, Maryland 21205, USA.
Yi
M
M
Ichikawa
N
N
LeBlond
G F
GF
Antonarakis
S E
SE
Ts'o
P O
PO
D001483:Base Sequence; D015139:Blotting, Southern / Q000379:methods*; D015342:DNA Probes; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D014644:Genetic Variation; D015894:Genome, Human; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid
D015342:DNA Probes