DNA was extracted from 300 µl of plasma using commercial DNA/RNA extraction kits (Qiagen, USA). The presence of HBV-DNA was confirmed and quantified using a previously described method [14] (link). For the preliminary genotyping, samples with detectable viremia were subjected to a nested PCR using a high proof reading enzyme mixture (Platinum PCR Super Mix High Fidelity, LifeTech, USA), that yielded a 879 bp fragment. Sequencing was performed in an automated DNA sequencer (ABI 3130xl, LifeTech, USA) using primers previously described [13] (link).
The full-length genome of Panamanian sequences clustering in particular clades of genotype F, were amplified using two protocols previously published [15] (link), [16] (link). When a sample failed to yield a good quality product with the one-step protocol, the nested approach was used.
The sequences obtained in this study were deposited in Genbank under the accession numbers KJ638656-KJ638679.
The full-length genome of Panamanian sequences clustering in particular clades of genotype F, were amplified using two protocols previously published [15] (link), [16] (link). When a sample failed to yield a good quality product with the one-step protocol, the nested approach was used.
The sequences obtained in this study were deposited in Genbank under the accession numbers KJ638656-KJ638679.
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