Her2 fish pharmdx kit

Example 2

This example compares the signal intensity and background from DNA probes and PNA probes on FFPE tissue sections using different solvents, at a denaturation temperature of 67° C. for 10 min and hybridization at 45° C. for 60 min.

FISH Probe Composition I: 3.3 ng/μL HER2 TxRed labeled DNA probe (⅓ of standard concentration) (size 218 kb) and ½ of the standard concentration (300 nM) of CEN17 FITC labeled PNA probes (both probes identical with probes from HER2 FISH pharmDx™ kit (K5331, Dako)); 15% ethylene carbonate (E26258, Sigma-Aldrich); 20% dextran sulfate; 600 mM NaCl; 10 mM citrate buffer, pH 6.2

FISH Probe Composition II: 3.3 ng/μL HER2 TxRed labeled DNA probe (⅓ of standard concentration) (size 218 kb) and ½ of the standard concentration (300 nM) of CEN17 FITC labeled PNA probes (both probes identical with probes from HER2 FISH pharmDx™ kit (K5331, Dako)); 15% 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (251569, Sigma-Aldrich); 20% dextran sulfate; 600 mM NaCl; 10 mM citrate buffer, pH 6.2

The scoring was performed on the mamma-carcinoma tissue of a multi-tissue section. Composition II was two-phased at the used composition concentrations.

Example 4

This example compares the signal intensity and background from DNA probes and PNA probes on FFPE tissue sections using DMSO, at a denaturation temperature of 67° C. for 10 min and hybridization at 45° C. for 60 min.

FISH Probe Composition I: 3.3 ng/μL HER2 TxRed labeled DNA probe (⅓ of standard concentration) (size 218 kb) and ½ of the standard concentration (300 nM) of CEN17 FITC labeled PNA probes (both probes identical with probes from HER2 FISH pharmDx™ kit (K5331, Dako)); 15% ethylene carbonate (E2625-8, Sigma-Aldrich); 20% dextran sulfate; 600 mM NaCl; 10 mM citrate buffer, pH 6.2

FISH Probe Composition II: 3.3 ng/μL HER2 TxRed labeled DNA probe (⅓ of standard concentration) (size 218 kb) and ½ of the standard concentration (300 nM) of CEN17 FITC labeled PNA probes (both probes identical with probes from HER2 FISH pharmDx™ kit (K5331, Dako)); 15% DMSO (Sigma-Aldrich); 20% dextran sulfate; 600 mM NaCl; 10 mM citrate buffer, pH 6.2

The scoring was performed on the mamma-carcinoma tissue of a multi-tissue section. Composition II with DMSO was unclear in appearance (milky white) at the used composition concentrations.

Example 3

This example compares the signal intensity and background from DNA probes and PNA probes on FFPE tissue sections using two non-cyclic solvents, at a denaturation temperature of 67° C. for 10 min and hybridization at 45° C. for 60 min.

FISH Probe Composition I: 3.3 ng/μL HER2 TxRed labeled DNA probe (⅓ of standard concentration) (size 218 kb) and ½ of the standard concentration (300 nM) of CEN17 FITC labeled PNA probes (both probes identical with probes from HER2 FISH pharmDx™ kit (K5331, Dako)); 15% N,N-dimethyl-acetamide (72336, Sigma-Aldrich); 20% dextran sulfate; 600 mM NaCl; 10 mM citrate buffer, pH 6.2

FISH Probe Composition II: 3.3 ng/μL HER2 TxRed labeled DNA probe (⅓ of standard concentration) (size 218 kb) and ½ of the standard concentration (300 nM) of CEN17 FITC labeled PNA probes (both probes identical with probes from HER2 FISH pharmDx™ kit (K5331, Dako)); 15% isobutyramide (144436, Sigma-Aldrich); 20% dextran sulfate; 600 mM NaCl; 10 mM citrate buffer, pH 6.2

The scoring was performed on the mamma-carcinoma tissue of a multi-tissue section. Composition I and II were two phased at the used composition concentrations.