Preparation of fluorescent in situ hybridisation probes without the need for optimisation of fragmentation
Patrick J McCoy, Anthony J Costello, Niall M Corcoran, Christopher M Hovens, Michael J Clarkson
MethodsX | ELSEVIER | Published : 2019
DNA-fluorescence in situ hybridisation (DNA-FISH) allows visualisation of chromosome organisation and rearrangement. FISH probes are pools of short fluorescently labelled DNA fragments that are often produced from template plasmids that contain large genomic inserts. For effective sample penetration and target hybridisation it is critical that probe fragments are between 200 and 500bp. Production of these short probes requires significant optimisation and can be confounded access to expensive sonication equipment or inherent sequence features that influence enzymatic fragmentation or amplification. Here we demonstrate that effective FISH probes can be prepared without the need for optimisati..View full abstract
Awarded by NHMRC
NMC is supported by a Movember - Distinguished Gentleman's Ride Clinician Scientist Award through Prostate Cancer Foundation of Australia's Research Program. This work was supported by NHMRC project grants 1104010 (C.M.H., A.J.C., N.M.C.) and 1047581 (C.M.H., A.J.C., N.M.C.), as well as a federal grant from the Australian Department of Health and Ageing to the Epworth Cancer Centre, Epworth Hospital (A.J.C., N.M.C., C.M.H. P.J.M, M.J.C). In carrying out this research, we received funding and support from Australian Prostate Cancer Research and the University of Melbourne, Australia.