HBV Transcripts Analysis With Northern Blot

HBV transcripts were analyzed by northern blot as previously described (27). Briefly, HBV RNA from transfected cells was extracted and purified with Trizol Reagent (Invitrogen) according to manufacturer instructions. The 4 µg from purified RNA were resolved on a 1.5% MOPS agarose gel containing 2.2 M formaldehyde, transferred onto a nylon membrane (GE Healthcare), immobilized with UV crosslinking, HBV transcripts corresponding to nucleotides 1072-2171 of HBV genome were detected with a DIG-labeled RNA probe. Dig Northern Starter Kit (Roche Diagnostics) was used for the preparation of probe and membrane detection. The quantity of 18S and 28S rRNAs were used as internal loading controls.