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Bacterial scRNA-seq library was prepared according to the protocol of VITAPilote kit (M20 Genomics, Hangzhou, China). In situ reverse transcription of bacteria was performed with random primers and the resulting cDNA fragment was added with adaptor. The droplet barcoding for a single bacterial cell was performed on VITACruiser Single Cell Partitioning System (M20 Genomics, Hangzhou, China). Bacterial cells, DNA extension reaction mix, and hydrogel barcoded beads were encapsulated using the VITACruiser. The aqueous phase containing cDNAs was purified with magnetic beads. The cDNAs were amplified by PCR, and purified with magnetic beads. All products were pooled to construct a standard sequencing library. Sequencing was done on a PE150 Illumina platform. |
RNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
PCR |
PAIRED
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