FISH bei Zytospins

  • Zytospins herstellen
  • Ca. 20 Minuten lang in einer Lösung aus 3:1 Methanol und Eisessig fixieren.
  • Jeweils 2 Minuten lang in 70 %, 85 % und 100 % Ethanol dehydrieren.
  • Die Objektträger an der Luft trocknen lassen.
  • 40 µl RNAse-Arbeitslösung (100 &mgr;g/ml) mit 40 ml vorgewärmtem (37 °C) 2x SSC mischen.
  • Objektträger ca. 15 Minuten bis 1 Stunde (je nach Zeitbeschränkungen) lang in die 37 °C warme RNase-Lösung legen
  • Die Objektträger dreimal jemals zwei Minuten lang in 2x SSC spülen.
  • Jeweils 2 Minuten lang in 70 %, 85 % und 100 % Ethanol dehydrieren. Bei Raumtemperatur trocknen lassen. Gemäß Packungsbeilage vorbereiten.

 Der RNAse-Schritt kann bei Zeitmangel weggelassen werden.

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