GSM8586483: Vibrio algalinolyticus biofilm scRNA-seq; Vibrio alginolyticus; RNA-Seq
The prepared bacteria suspension was permaeablized and blocked for single-stranded DNA-blocking. Random primers were then added for the reverse transcription of total RNA inside the cell. Subsequently, the generated cDNA fragments were ligated to adaptors inside the bacteria. The reverse-transcribed single-bacteria suspension and reagents, along with barcode beads containing cell barcodes and UMIs, were mixed. The mixture underwent encapsulation, capturing and barcoding using the VITAcruizer DP400 instrument. The resulting product underwent extension, resulting in barcoded cDNA strands. PCR amplification was conducted using the cDNA as the template. The purified products after cDNA amplification were then used to constructed a standard next-generation sequencing library. The constructed single-cell library contained P5 and P7 adapters and was sequenced on a Xplus sequencer (Illumina) with 150 bp paired-end reads.