INTRODUCTION

BioSpyder is offering a proprietary high-throughput single-cell genomics platform, TempO-LINC (Ligation INdexed Cells) . This product relies on the foundational TempO-Seq technology approach developed by BioSpyder for gene expression analysis. The reliance of TempO-LINC on the TempO-Seq technology makes it a fundamentally different and novel approach to single-cell genomics. Instead of reverse transcribing RNA, TempO-Seq uses detector oligos (DOs) to specifically hybridize to two immediately adjacent regions within an expressed transcript. The two DOs hybridized to each transcript are then ligated together forming a DNA product that can be amplified and sequenced, thereby enabling the specific and sensitive detection of gene expression. This approach has many advantages including the ability to either be targeted to small numbers of transcripts or highly multiplexed across the entire transcriptome, reduced read mapping and bioinformatics challenges, reduced sequencing costs and the ability to assay gene expression on fixed cells that are banked in the freezer.

TEMPO-LINC WORKFLOW

APPROACH

TempO-LINC relies on the exact same DO pools that BioSpyder currently sells for whole transcriptome analysis; however, once ligated within fixed cells, the DO product is barcoded through multi-step barcoding which enables unique identifiers to be generated for each cell. Following our proprietary barcoding step, samples are PCR amplified and purified without the need for template switch synthesis, fragmentation, or SPRI selection steps. Unlike droplet-based methods which ultimately have finite limits on microfluidic processing and sample volumes, TempO-LINC is a truly scalable approach that is limited only by the number of barcodes in each round and the number of rounds of barcoding.