Replication data for: A novel sweet potato potyvirus open reading frame (ORF) is expressed via polymerase slippage and suppresses RNA silencing

Total RNA was extracted with TRIzol LS Reagent (Invitrogen) according to the manufacturer’s recommendations alow-molecular-weight (LMW) and high-molecular-weight (HMW) RNA fractions were separated as described in Cuellar et al. (2009).

LMW RNA (15 lg/lane) and HMW RNA (10 lg/lane) were separated by electrophoresis in a denaturing 15% polyacrylamide gel and 1.2% (w/v) agarose gel, respectively, both containing formaldehyde. The two gels were transferred to a nylon membrane (GE Healthcare) and crosslinked chemically with 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide (Pall and Hamilton, 2008) or by UV light (Sambrook and Russell, 2001), respectively. A GFP mRNA-specific radioactive RNA probe was synthesized, fractionated with carbonate hydrolysis and used to probe the membranes for GFP mRNA and GFP mRNA-derived siRNA, as described previously (Cuellar et al., 2009).