Treatments of 0.5 μg of the total genomic DNA with RNase HI (Takara) or RNaseHII (NEB) were performed as described (18 (link)). It was critical to run the reaction samples on the gel followed up by electric transfer to separate RNase HI from the substrate. The enzyme tightly binds to the substrate (even at 0°C in water) and completely blocks immunodetection if a reaction mix is directly used in the dot-blot procedure.
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