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Parallel genome analysis by one‐ and two‐dimensional DNA fingerprinting in human gliomas
Author(s) -
Nürnberg Peter,
Marczinek Karola,
Thiel Gundula,
Hampe Jochen
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501601284
Subject(s) - microsatellite , biology , dna , dna profiling , microbiology and biotechnology , genomic dna , dna sequencing , oligonucleotide , carcinogenesis , genome , genetics , gene , allele
The detection of DNA variation in cancers is an important step in elucidating the mechanism of tumorigenesis. Using the strategy of multipoint genome analysis we detected many differences between glioma‐derived and constitutional DNA by customary DNA fingerprinting with simple repetitive oligonucleotide probes. Amplification of the epidermal growth factor receptor (EGFR) gene has been found to be easily detectable as new or highly intensified bands in one‐dimensional (1‐D) DNA fingerprints of glioblastoma DNA generated with probes (GTG) 5 or (GT) 8 . However, in most low‐grade astrocytomas, 1‐D DNA fingerprinting has failed to reveal any genomic abnormalities. In these cases a two‐dimensional (2‐D) technique was successfully employed that is based on size separation in neutral gels followed by sequence‐dependent separation in denaturing gradient gels and hybridization with several mini‐ and microsatellite core probes. The hundreds of spots visualized with this technique were used to detect subtle changes probably occurring as the initial steps of tumorigenesis in human gliomas. On average, five of the approximately 580 sports generated by probes CAC and 33.6 were found to be altered in tumor DNA; 80% of the alterations were spot losses, the rest being spot gains or amplifications. Computer‐based image analysis using an external lambda marker provided a stringent way to compare spot patterns generated by 2‐D DNA finger‐printing. In comparisons performed between typing patterns generated on the same gel, 99% of truly identical spots were confirmed by the sofware. In intergel comparisons 84% of identical spots were matched on the basis of the marker information alone.