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Title

Two-dimensional electrophoresis southern transfer method for detecting human genome variability using a LINE-1 sequence probe

Authors
Nakashima, H.
Yi, M.
Ichikawa, N.
LeBlond, G. F.
Ts'o, P. O.
Published in Analytical Biochemistry. 1995, vol. 227, no. 2, p. 319-327
Abstract 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.
Keywords Base SequenceBlotting, Southern/ methodsDNA ProbesElectrophoresis, Gel, Two-DimensionalFemaleGenetic VariationGenome, HumanHumansMaleMolecular Sequence DataRepetitive Sequences, Nucleic Acid
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PMID: 7573953
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NAKASHIMA, H. et al. Two-dimensional electrophoresis southern transfer method for detecting human genome variability using a LINE-1 sequence probe. In: Analytical Biochemistry, 1995, vol. 227, n° 2, p. 319-327. https://archive-ouverte.unige.ch/unige:8919

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Deposited on : 2010-07-12

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