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Labeling Vg with DIG was performed according to the instruction of DIG protein labeling kit (Roche).
In situ hybridization with isotopes or digoxygenin (DIG) was performed as previously described.
Visualization of DIG was performed by incubation with a monoclonal antibody coupled with alkaline phosphatase (anti-DIG alkaline phosphatase Fab fragments diluted 1 500; Boehringer Mannheim) at 20°C for 2 h.
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The group also hosts fossil digs where excavation is performed in the context of Noah and the biblical flood.
After blocking the slides, immunological detection for DIG with anti-DIG-AP was performed according to the instruction manual of the DIG Nucleic Acid Detection Kit (Roche).
Detection using anti-dig-rhodamine was performed according to [ 37].
As extensive transcriptional profiling has already been reported for incompatible A. brassicicola- Arabidopsis interaction (incompatible), an initial study of the A. brassicicola-DiG pathosystem was performed.
The hybridization of the DIG-labelled probes was performed according to the DIG user's manual (www.roche-applied-science.com).com
Immunological detection of DIG-labeled hybrids was performed with an anti-DIG antibody from sheep conjugated with alkaline phosphatase as recommended by the manufacturer (Roche).
Detection of the DIG-labeled probe was performed with alkaline phosphatase-conjugated anti-DIG antibody using a HNPP Fluorescence Detection kit (Roche Diagnostics GmbH).
Detection of binding of Dig-conjugated antibodies was performed using HRP-conjugated sheep anti-Dig Abs (Fab fragments, Roche, Mannheim, Germany).
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