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BRIGITTE SCHLEGELBERGER et aI.
Hybridization mixture for indirectly labeled centromeric probes (Oncor):
Add 1 III of biotin labeled probe (Oncor Cat. No. P5000-BIO -P5090-BIO)
or 1 III of digoxigenin labeled probe (Oncor Cat. No. P5000-DG-P5090-DG)
or 0.5 III of biotin labeled probe (Oncor Cat. No. P5000-BIO -P5090-BIO)
and 0.5 III of digoxigenin labeled probe (Oncor Cat. No. P5000-DG-P5090DG) as supplied by the manufacturer to 9 III of "mastermix for centromeric probes". Non-commercial DNA probes should be adjusted to 10
ng/Ill in TE buffer (l mM EDTA, 10 mM Tris-HCl, pH 8) before being
added to the mastermix.
Hybridization mixture for indirectly labeled commercial single copy
probes (Oncor):
Indirectly labeled single copy probes (Oncor), e.g. probes for translocations or microdeletions, can be applied without prior modifications.
For the combination of single copy probes with centromeric probes a
10:1 mixture, for the combination of two single copy probes a 1:1 mixture
is recommended. If hybridization with the directly applied single copy
probe or a mixture of two probes produces a high background noise a
1:2 or 1:4 (up to 1:10) dilution of the probe/probe mixture in "mastermix
for single copy probes plus Cot-1 DNA" may enhance the hybridization
quality.
Hybridization mixture for indirectly labeled non-commercial single copy
probes with or without combination with indirectly labeled centromeric
probes (Oncor):
For non-commercial single-copy probes the mixture of 1 III labeled singlecopy probe (40-100 ng/Ill) and 9 III "mastermix for single copy probes plus
Cot-1 DNA" is recommended. If the probe produces a high background
noise or results in a low signal intensity 1-5 III labeled single-copy probe
can be coprecipitated with 100 III Cot-1 DNA using conventional ethanol
precipitation (as described for "mastermix for single copy probes plus
Cot-1 DNA") and resuspended in 10 III "mastermix for single copy
probes". For combining a single-copy probe with a centromeric probe
(Oncor) add 1 III of the latter to the respective probe mixture.
BRIGITTE SCHLEGELBERGER et aI.
Hybridization mixture for indirectly labeled centromeric probes (Oncor):
Add 1 III of biotin labeled probe (Oncor Cat. No. P5000-BIO -P5090-BIO)
or 1 III of digoxigenin labeled probe (Oncor Cat. No. P5000-DG-P5090-DG)
or 0.5 III of biotin labeled probe (Oncor Cat. No. P5000-BIO -P5090-BIO)
and 0.5 III of digoxigenin labeled probe (Oncor Cat. No. P5000-DG-P5090DG) as supplied by the manufacturer to 9 III of "mastermix for centromeric probes". Non-commercial DNA probes should be adjusted to 10
ng/Ill in TE buffer (l mM EDTA, 10 mM Tris-HCl, pH 8) before being
added to the mastermix.
Hybridization mixture for indirectly labeled commercial single copy
probes (Oncor):
Indirectly labeled single copy probes (Oncor), e.g. probes for translocations or microdeletions, can be applied without prior modifications.
For the combination of single copy probes with centromeric probes a
10:1 mixture, for the combination of two single copy probes a 1:1 mixture
is recommended. If hybridization with the directly applied single copy
probe or a mixture of two probes produces a high background noise a
1:2 or 1:4 (up to 1:10) dilution of the probe/probe mixture in "mastermix
for single copy probes plus Cot-1 DNA" may enhance the hybridization
quality.
Hybridization mixture for indirectly labeled non-commercial single copy
probes with or without combination with indirectly labeled centromeric
probes (Oncor):
For non-commercial single-copy probes the mixture of 1 III labeled singlecopy probe (40-100 ng/Ill) and 9 III "mastermix for single copy probes plus
Cot-1 DNA" is recommended. If the probe produces a high background
noise or results in a low signal intensity 1-5 III labeled single-copy probe
can be coprecipitated with 100 III Cot-1 DNA using conventional ethanol
precipitation (as described for "mastermix for single copy probes plus
Cot-1 DNA") and resuspended in 10 III "mastermix for single copy
probes". For combining a single-copy probe with a centromeric probe
(Oncor) add 1 III of the latter to the respective probe mixture.