Genomic DNA (gDNA) was extracted from cells and 20 mg fresh liver tumor tissues using the Genomic DNA Miniprep Kit (AxyGEN, USA) according to the manufacturer’s instructions. As done previously [44 (link)], 35 targeted sites of gDNA and eccDNAs were amplified by PCR with primers listed (Additional file 1 : Table S1). PCR products were purified by AxyPrep DNA Gel Extraction Kit or AxyPrep PCR Cleanup Kit (AxyGEN). Purified PCR products of each target were combined into a tube with equal volume. One microgram of PCR product mixture was used to establish DNA fragment libraries by VAHTS Universal DNA Library Prep Kit for Illumina V2 (Vazyme Biotech, China) and sequenced on Illumina HiSeq 2500.
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