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Finally, we normalized raw counts into counts per million (CPM), computed the differential expression analysis between the two conditions of the experimental design, and annotated each shRNA with the corresponding gene

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For signal detection, the ABI Prism 7700 sequence detector (Life Technologies, CA, USA) was programmed to execute an initial step of 2 min at 50C and 10 min at 95C, followed by 40 thermal cycles of 15 sec at 95C and 1 min at 60C