High resolution mapping using fluorescence in situ hybridization to extended DNA fibers prepared from agarose-embedded cells
Heiskanen, M.; Karhu, R.; Hellsten, E.; Peltonen, L.; Kallioniemi, O.P.; Palotie, A.
Biotechniques 17(5): 928
1994
ISSN/ISBN: 0736-6205 PMID: 7840975 Document Number: 437999
Fluorescence in situ hybridization (FISH) and pulse field gel electrophoresis (PFGE) are essential techniques in physical mapping and in positional cloning. We present a technique that utilizes agarose-embedded high molecular weight DNA prepared for PFGE as a target for FISH. The agarose blocks are melted, and the DNA is extended on a poly-L-lysine-coated microscope slide. The resulting DNA fibers appear on the slide as long straight strands and are a suitable target for high resolution FISH mapping as demonstrated here with cosmid and plasmid hybridizations.