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BRIGITTE SCHLEGELBERGER et al.
- DABCO (l,4-Diazabicyclo-(2,2,2)-octane) (Sigma D2522)
- DAPI (4,6-Diamidino-2-phenylindole) (Sigma D9542)
- Deionized formamide (Gibco BRLlLife Technologies 15515-018)
- Dextran sulfate 50% (Q-BIOgene DEXT 5001)
- Digoxigenin-conjugated sheep anti-mouse antibody (Chemicon International AQ 300D)
- FITC-conjugated donkey anti-mouse antibody (Jackson 715-095-151)
- FITC-conjugated sheep anti-digoxigenin antibody (Roche 1207741)
- Mouse anti-FITC antibody (monoclonal) (Dakopatts M878)
- Mouse anti-digoxigenin antibody (monoclonal) (Roche 1333062)
- Normal mouse serum (Jackson 015-000-001)
- Rubber cement: Fixogum (Marabu 2901 10000)
Buffers and
solutions
- Antifade solution: To 90 ml glycerol add 10 ml PN buffer containing
23 mg/ml DABCO
- DAPI stock solution: 0.2 mg/ml DAPI in A. bidest
- Paraformaldehyde solution (1%): Dissolve 1 g paraformaldehyde powder (harmful!) in 60 ml A. bidest.; add 5 drops 10 M NaOH. Heat to 70100°C until the solution is clear when viewed from above the bench.
Cool down and add 10 ml100 mM MgCI 2 • Adjust pH to 7-7.5 and fIll up
to 100 ml with distilled water. Filter the solution. Store at 4°C.
- PN buffer: 0.1 M NaH z P0 4 , 0.1 M Na z HP0 4 , pH 8.0
- PNM buffer: Add 5% dry milk powder to PN buffer; heat to 50°C overnight while stirring; add 0.03% NaN 3 (poisonous); PNM buffer is stable
at 4°C for at least 6 months.
- 20xSSC: x3 M NaCl, 0.3 M sodium citrate; pH 7.0
- Mastermix for indirect centromeric probes:
Mix 6 ml deionized formamide, 2 ml 50% dextran sulfate and 0.5 ml
20xSSC. Vortex thoroughly, adjust to pH 7.5 and make up to a final
volume of 9 ml with A.bidest.
BRIGITTE SCHLEGELBERGER et al.
- DABCO (l,4-Diazabicyclo-(2,2,2)-octane) (Sigma D2522)
- DAPI (4,6-Diamidino-2-phenylindole) (Sigma D9542)
- Deionized formamide (Gibco BRLlLife Technologies 15515-018)
- Dextran sulfate 50% (Q-BIOgene DEXT 5001)
- Digoxigenin-conjugated sheep anti-mouse antibody (Chemicon International AQ 300D)
- FITC-conjugated donkey anti-mouse antibody (Jackson 715-095-151)
- FITC-conjugated sheep anti-digoxigenin antibody (Roche 1207741)
- Mouse anti-FITC antibody (monoclonal) (Dakopatts M878)
- Mouse anti-digoxigenin antibody (monoclonal) (Roche 1333062)
- Normal mouse serum (Jackson 015-000-001)
- Rubber cement: Fixogum (Marabu 2901 10000)
Buffers and
solutions
- Antifade solution: To 90 ml glycerol add 10 ml PN buffer containing
23 mg/ml DABCO
- DAPI stock solution: 0.2 mg/ml DAPI in A. bidest
- Paraformaldehyde solution (1%): Dissolve 1 g paraformaldehyde powder (harmful!) in 60 ml A. bidest.; add 5 drops 10 M NaOH. Heat to 70100°C until the solution is clear when viewed from above the bench.
Cool down and add 10 ml100 mM MgCI 2 • Adjust pH to 7-7.5 and fIll up
to 100 ml with distilled water. Filter the solution. Store at 4°C.
- PN buffer: 0.1 M NaH z P0 4 , 0.1 M Na z HP0 4 , pH 8.0
- PNM buffer: Add 5% dry milk powder to PN buffer; heat to 50°C overnight while stirring; add 0.03% NaN 3 (poisonous); PNM buffer is stable
at 4°C for at least 6 months.
- 20xSSC: x3 M NaCl, 0.3 M sodium citrate; pH 7.0
- Mastermix for indirect centromeric probes:
Mix 6 ml deionized formamide, 2 ml 50% dextran sulfate and 0.5 ml
20xSSC. Vortex thoroughly, adjust to pH 7.5 and make up to a final
volume of 9 ml with A.bidest.