Whole library-amplification and labelling of human chromosome-specific composite probes for fluorescence in situ hybridization (FISH) using PCR.
Chromosome painting with human-specific, whole-plasmid library probes has become a widespread technique in cytogenetic research and radiation biology. Fast and convenient methods for probe amplification and labelling are required that conserve the complexity of the libraries and stain the entire length of selected chromosomes. The present study uses PCR for amplification and labelling of the whole bluescribe plasmid libraries pBS1, pBS4 and pBS12 using M13 forward and reverse sequencing primer. Amplification and labelling can be accomplished within a few hours with a minimum of laboratory equipment and costs. It is therefore suitable for all laboratories that do not have the facilities for growing transformed bacteria containing plasmids with inserts of human origin.
Oberheitmann
B
B
Department of Environmental Epidemiology, University of Bremen, Germany.
D002869:Chromosome Aberrations; D015342:DNA Probes; D015723:Gene Library; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D016133:Polymerase Chain Reaction
D015342:DNA Probes