RT Journal Article
SR Electronic
T1 Improved in situ detection method for telomeric tandem repeats in metaphase spreads and interphase nuclei
JF Molecular Pathology
JO Mol Pathol
FD BMJ Publishing Group Ltd and Association of Clinical Pathologists
SP 48
OP 50
DO 10.1136/mp.53.1.48
VO 53
IS 1
A1 V Uhlmann
A1 M Prasad
A1 I Silva
A1 K Luettich
A1 L Grande
A1 L Alonso
A1 M Thisted
A1 K J Pluzek
A1 J Gorst
A1 M Ring
A1 M Sweeney
A1 C Kenny
A1 C Martin
A1 J Russell
A1 N Bermingham
A1 M O'Donovan
A1 O Sheils
A1 J J O'Leary
YR 2000
UL http://mp.bmj.com/content/53/1/48.abstract
AB Peptide nucleic acid technology (PNA) has become an extremely useful tool and promises to impact on molecular biology and diagnostics. These synthetic DNA analogues pair with DNA and RNA molecules according to Watson and Crick base pairing rules. This paper describes a sensitive and quick fluorescent in situ hybridisation (ISH) technique to determine DNA telomere repeat sequences (TTA GGG)n using epifluorescence microscopy. Telomeres are special, repeated structures at the end of each eukaryotic chromosome and serve as protective caps to prevent DNA rearrangements and fusion of chromosomes. A model system has been developed, using stimulated peripheral blood lymphocytes, which facilitates simultaneous detection of telomeres in metaphase as well as in interphase nuclei. A fluorescein isothiocyanate labelled PNA probe (18 mer) directed against complementary telomeric sequences at the end of each chromosome is used. In addition, a simple, easy to perform PNA-ISH protocol is described that overcomes common hybridisation problems encountered using DNA and RNA oligoprobes. Furthermore, the usefulness of a chromogenic immunocytochemical detection system is shown for PNA-ISH.