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Assignment of Alu-repetitive sequences to large restriction fragments from human chromosomes 6 and 22
Authors:Eckart Meese  Hans-Werner Müller  Nicole Brass  Jeffrey M. Trent  Nikolaus Blin
Affiliation:(1) Medical School, Inst. of Human Genetics, Bau 68, 66421 Homburg/Saar, Germany;(2) National Center for Human Genome Research, NIH, Bldg. 49, Rm 4A, 20892 Bethesda, MD, USA;(3) Div. of Molecular Genetics, University of Tübingen, Wilhelmstr. 27, 72074 Tübingen, Germany
Abstract:
We have employed a pulsed field gel electrophoresis and Alu hybridization approach for identification of large restriction fragments on chromosome 6 and 22. This technique allows large portions of selected human chromosomes to be visualized as discrete hybridization signals. Somatic cell hybrid DNA which contains chromosome 6 or chromosome 22 was restricted with either Notl or Mlul. The restriction fragments were separated by pulsed field gel electrophoresis (PFGE) and hybridized against an Alu repetitive sequence (Blur 8). The hybridization signals result in a fingerprint-like pattern which is unique for each chromosome and each restriction enzyme. In addition, a continuous pattern of restriction fragments was demonstrated by gradually increasing puls times. This approach will also be suitable to analyze aberrant human chromosomes retained in somatic cell hybrids and can be used to analyze flow sorted human chromosomes. To this end, our method provides a valuable alternative to standard cytogenetic analysis.
Keywords:PFGE (pulsed field gel electrophoresis)  repetitive DNA  chromosomal sequences  mapping
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