ASV-Seq: HepG2 cell line
Target sequencing was performed with the assistance of TinyGen Company. Firstly, the sample was prepared for library construction using specific primers. The library construction involved a two-step PCR amplification method. In the first step, the target fragments were amplified using specific primers, and the amplified fragments were then subjected to gel purification. Subsequently, the purified products were used as templates for a second round of PCR amplification, to add the Illumina platform-required adaptors, sequence primers, and barcodes to both ends of the target fragments. Afterward, Illumina high-throughput sequencing was performed. The obtained paired-end reads were first sorted based on barcodes to distinguish each sample. Then, the sequences underwent quality control and filtering. They were assembled based on overlap relationships, and the assembled sequences underwent another round of quality control and filtering to obtain optimized sequences. The optimized sequences were subjected to ASV (amplicon sequence variant) analysis and taxonomic classification. Based on the ASV analysis results, a table was generated that showed the distribution of ASVs for each sample. The total count of sequences that matched the plasmid sequences at both ends was used as the denominator, and the count of sequences with different mutation types in each sample was used as the divisor to calculate the mutation rate.
SUB14660169
TLS-A3