Probe-based capture sequencing of Hepatitis B Virus: children liver
For successful sequencing of the hepatitis B virus virus genome with low numbers of viral copies, we used the in-solution target enrichment strategy . Library preparation was conducted using IGT Enzyme Plus Library Prep Kit V3 and IGT Adapter & UDI Primer (iGeneTech) to generate dual-indexed libraries. Hybridization of DNA baits with capture pools was performed at 50 C for 18h. Theprobes of hepatitis B virus was designed by iGeneTech. Dynabeads MyOne Streptavidin T1 magnetic beads (Life Technologies) were used to isolate biotinylated DNA, and amplification of bead-bound enriched libraries (18-22 cycles) was performed using Post PCR Master Mix and Post PCR Primer (iGeneTech). We used the Trimmomatic software to perform quality control on the rawdata, remove adapters and low-quality data, and obtain cleandata. The MEGAHIT software was used to assemble the aligned reads, obtaining the total contig sequence. Use the blastn software to align the contig sequence with the human genome, remove the human-derived contigs, obtain the non-human contig sequence, and then align it with the microorganism genome sequence to obtain the microbial typing results. Finally, we determined the final consensus sequence on the basis of the reference genome sequence.