Single-cell Sequencing Immune Landscape of Hepatocellular Carcinoma after Transarterial Chemoembolization
The indexed sequencing libraries of each sample was processed independently for (a) fragmentation, end repair and A-tailing; (b) size selection with SPRI select beads; (c) adaptor ligation; (d) post-ligation cleanup with SPRI select beads; and (e) sample index PCR and final cleanup with SPRI select beads. The final single cell sequencing library comprises the standard Illumina paired-end constructs which begin and end with P5 and P7 primers used in Illumina bridge amplification. The barcoded sequencing libraries were quantified by a Bioanalyzer Agilent 2100 System using a High Sensitivity DNA chip (Agilent), and the quantitative PCR using a KAPA Library Quantification Kit (KAPA Biosystems). Finally, the sequencing libraries were loaded onto Novaseq 6000 (Illumina) platform to generate 2 150-bp paired-end reads.