For qualified libraries, we performed paired-end sequencing (2250 bp) using the NovaSeq 6000 SP Reagent Kit (500 cycles) on the Illumina NovaSeq platform. First, the libraries to be sequenced (with non-repeating indexes) were gradient-diluted to 2 nM and then pooled according to the required data volume ratio. The pooled libraries were denatured into single strands using 0.1N NaOH before loading onto the sequencer. The amount of loaded libraries could be adjusted between 15-28 pM based on actual conditions.