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For interphase preparations no thymidine or Colcemid were used.
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Interphase preparations for MN assay and fluorescence in situ incorporation were obtained following the procedures previously described [ 1- 4, 6, 23].
Single-layer nuclei preparations for interphase FISH hybridisations were prepared using the Hedley method with modifications (Castro et al, 1993).
For light microscopy, BF-fixed nuclei thus provide a gold standard for interphase FISH against which other preparation methods can be compared.
He is a manufacturer's representative in Tampa, Fla., for Interphase Technologies, a marine electronics concern in Santa Cruz, Calif.
However, for interphase analysis, samples were produced without colcemid treatment.
Interphase cell preparations were incubated at 37°C for 10 min and then treated with 300 μl ml−1 HCl pepsin, pH 2.7 3 (Sigma, Dorset, UK) at 37°C, to remove cytoplasmic proteins, hence improving probe penetration.
This proximal pool comprises cells dividing mitotically, as well as cells completing their final passage through interphase in preparation for entry into the meiotic cell cycle.
In our study, the cut-off level for interphase-FISH was 5%.
Interphase nuclei preparation was performed directly from these cultures.
Fluorescence in situ hybridisation (FISH) has been used increasingly for gene mapping and ordering probes on interphase and metaphase preparations.
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