Figure 1

Tag-linkage sequencing of HBV RT/S region. (a) A schematic presentation of the HBV viral genome region (polymerase gene) that was processed and reconstructed using the tag-linkage method. 836 bp of polymerase gene (blue box) was recovered, including the majority of the reverse transcriptase domain (overlapping S gene) and part of the RNase H gene. (b) A schematic presentation of the experiment flow for tag linkage deep sequencing. 1) HBV sequences were amplified from viral DNA extracted from patient plasma using primers targeting conserved region. Tags were added onto each molecule for sequencing error correction and haplotype reconstruction. 2) Different length of sequences (covering different number of non-overlapping small amplicons) were amplified from ~10,000 molecules. 3) Amplified sequences were ligated. 4) Small amplicons were further amplified for sequencing.