Qubit ssdna analysis kit

scRNA sequencing libraries were constructed using DNelab C4 and following the manufacturer’s instructions (Liu et al., 2019 ). Libraries were quantified using a Qubit ssDNA Analysis Kit (Thermo Fisher Scientific) and sequenced using a DIPSEQ T1 sequencer from the China National Gene Bank. Briefly, single-cell suspensions were used for droplet generation, emulsion breakage, bead collection, reverse transcription, and cDNA amplification was used to generate barcoded libraries. Indexed single-cell RNA-seq libraries were constructed according to manufacturer’s protocol. Sequencing libraries were quantified using the QubitTM ssDNA Assay Kit (Thermo Fisher Scientific, #Q10212). DNA nanoballs were loaded into the patterned nanoarrays and sequenced on an ultra-high-throughput DIPSEQ T1 sequencer using the following read lengths: 30 bp for read 1 (inclusive of 10 bp cell barcode 1), 10 bp for cell barcode 2, 10 bp for the unique molecular identifier; 100 bp of transcript sequence for read 2, and 10 bp for the sample index.

ScRNA sequencing libraries were constructed using DNelabC4 following the manufacturer's instructions.⁶⁴ The libraries were quantified using a Qubit ssDNA analysis kit (Thermo Fisher Scientific) and sequenced using the DIPSEQ T1 sequencer of the China National Gene Bank (CNGB).