Cell capture was performed using the Single Cell 5' Library and Gel Bead Kit and the Chromium Single Cell A Chip Kit (10x Genomics, CA USA), according to the manufacturer's protocol. Captured cells were lysed and the released RNA was barcoded by reverse transcription in individual Gel Bead in Emulsions (GEMs). The complementary DNA (cDNA) was generated, amplified, and quality assessed. Subsequently, the scRNA-seq library was constructed, as previously described (27 (link),28 (link)). Finally, the library was sequenced using an Illumina Novaseq6000 sequencer. The sequencing depth was at least 100,000 reads per cell, using a pair-end 150 bp (PE150) reading strategy.
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Single-cell RNA sequencing protocol
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Corresponding Organization : Soochow University
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Variable analysis
independent variables
- Cell capture technique (using the Single Cell 5' Library and Gel Bead Kit and the Chromium Single Cell A Chip Kit from 10x Genomics)
- Sequencing depth (at least 100,000 reads per cell)
- Sequencing strategy (pair-end 150 bp)
- Manufacturer's protocol for cell capture, library construction, and sequencing
- No positive or negative controls were explicitly mentioned in the provided information.
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