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Scientific article
English

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

Published inAnalytical biochemistry, vol. 227, no. 2, p. 319-327
Publication date1995
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 Sequence
  • Blotting, Southern/ methods
  • DNA Probes
  • Electrophoresis, Gel, Two-Dimensional
  • Female
  • Genetic Variation
  • Genome, Human
  • Humans
  • Male
  • Molecular Sequence Data
  • Repetitive Sequences, Nucleic Acid
Citation (ISO format)
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. doi: 10.1006/abio.1995.1287
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ISSN of the journal0003-2697
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