Cloning and sequencing of PCR products
The RT-PCR products were purified and ligated into
pGEM T-easy vector (Promega, USA) and recombinant
plasmids from transformed cells were subjected
to restriction analysis to verify the presence of an insert
of the expected fragment. Cloned viroid cDNAs
were sequenced with a Beckman 2000XL sequencer.
Multiple alignment and sequence analysis were performed
using the BioEdit software (version 5.0.9,
available at http://www.mbio.ncsu.edu/BioEdit/bioedit.
html).