Single cell RNA-seq with ultra-high sensitivity and a streamlined workflow

The new plexWell Rapid Single Cell RNA Library Prep Kit uses an optimised chemistry for highly sensitive full-length cDNA synthesis. It captures the complexity of the transcriptome, including splice variants, fusion transcripts and SNPs.

The innovative tagmentation-based library preparation reduces hand-on time, removes the need for sample normalization and allows for highly multiplexed sequencing.

Outperformance vs. common scRNA-seq methods

The plexWell Rapid Single Cell RNA Library Prep Kit was benchmarked against common Smart-seq protocols (1,2) using a challenging sample with ultra-low cellular RNA content.

The plexWell Kit achieved the highest sensitivity (Fig.1). Furthermore, the plexWell Kit delivered a very uniform gene body coverage, the highest percentage of uniquely mapping reads, the highest exon- and the lowest intron-mapping rate.

Fig.1: Benchmarking the plexWell Rapid Single Cell RNA Library Prep Kit, and downsampling effects on gene detection. CD45+ human PBMCs were processed in 96-well plates according to the manufacturer’s instructions or published protocols (1,2).

Data from Simone Picelli, Single Cell Genomics Platform, IOB, Basel, Switzerland.

Streamlined library prep with built-in normalization

The plexWell Rapid Single Cell RNA Library Prep Kit facilitates library preparation with a unique barcoding scheme and built-in automatic normalization, reducing the need for sample QC. The whole process from sorted cells to sequencing-ready libraries can be completed in the same day, without any specialized equipment.

Fig.2: plexWell workflow vs. conventional library preparation.

Multiplexing uniformity

plexWell auto-normalization ensures a highly even read distribution across single cells. Different sets of barcodes are available for multiplexing up to 1152 cells in one sequencing run.

Fig.3: All 96 cDNA samples were quantified and normalized for the Smart-seq2 workflow. For the plexWell workflow, cDNAs were diluted, by applying a global dilution factor derived from QC data for only 24 of the samples.

References

  1. Picelli et al., Nature Protocols 9(1):171-81 (2014)
  2. Hagermann-Jenssen et al., Nature Biotechnology 38(6):708-714 (2020)

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