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FISH Procedure for FFPE Specimens

Reagents Required but Not Provided

·         Paraffin Pretreatment Reagent Kit (Cat No: CT-ACC112-05):

o    Pretreatment Solution (50 ml): store at room temperature (RT)

o    Protease Buffer (62.5 ml, pH 2.0): store at RT

o    Protease (250 mg): Lyophilized, store at -20°C

·         FISH Reagent Kit (Cat No: CT-ACC101-20):

o    20X Sodium Chloride-Sodium Citrate Buffer (SSC) Salt: store at RT, avoid humidity

o    4’,6-diamidino-2-phenylindole (DAPI) Counterstain: store at 4°C in the dark

o    NP-40 (octylphenoxypolyethoxyethanol, or Nonidet P-40): store at RT

·         Xylene: store at RT

·         Ethanol (100%): store at RT

·         Purified water: store at room RT

·         Concentrated (12N) HCl: store at room RT

 

Preparation of Working Solutions

1. 20X SSC Solution (pH 7.0)

Reagents

Amount added

Final Concentration

SSC Salt

66 g

20X

Deionized H2O (dH2O)

250 ml

 

TOTAL

250 ml

 

 

2. Protease Solution

Reagents

Amount added

Final Concentration

   

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