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For the directly labelled PNA probe, after stringent washes in 2 × SSC, 0·1 × SSC and 2 × SSC with 0·2% Tween-20 at 42 °C, for 5 min each, preparations were mounted in antifade buffer Vectashield (Vector Laboratories, Peterborough, UK) containing 4′,6-diamidino-2-phenylindole (DAPI) counterstain (2 μg mL−1), and stored at 4 °C.
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