FISH su citocentrifugati

  • Preparazione di citocentrifugati (cytospin)
  • Fissare in 3:1 metanolo:acido acetico glaciale per ~20 minuti
  • Disidratare in soluzioni di etanolo al 70%, 85% e 100% per 2 minuti in ciascuna
  • Far asciugare i vetrini all'aria
  • Mescolare 40 ul di soluzione di lavoro di RNAsi (100 ug/ml) con 40 ml di 2x SSC preriscaldata (37°C)
  • Collocare i vetrini nella soluzione di RNAsi a 37°C per ~15 minuti - 1 ora (a seconda dei limiti di tempo)
  • Sciacquare i vetrini in 2x SSC per tre volte, due minuti per volta
  • Disidratare in soluzioni di etanolo al 70%, 85% e 100% per 2 minuti in ciascuna. Far asciugare a temperatura ambiente. Preparare in base a quanto riportato nel foglietto illustrativo.

 Il passaggio relativo alla RNAsi può essere saltato se il tempo non lo consente.

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