TempO-Seq workflow

Based on BioSpyder Technologies’ proprietary Templated Oligo Detection Assay, TempO-Seq can quantitate targeted transcripts in an easy to follow workflow that does not require dedicated equipment. It can be run in a standard PCR instrument or microplate incubator manually or using standard pipetting platforms. The assay is highly amenable to automation, enabling implementation on 96-, 384-, and 1536-well formats. Sample barcoding, together with sequencing of short templates to measure each gene, allows pooling up to 6,144 samples in one sequencing run. Assay content is flexible and customisable, from focused panels monitoring specific genes or cellular pathways up to the whole transcriptome, delivering unprecedented accuracy and sensitivity for low level inputs. You can select focused content from an archive of detector oligos measuring the whole transcriptome, and add additional custom content such as the measurement of specific isotypes, fusions, or mutations. Together with robust probe design and simplified data analysis that eliminates the need for bioinformatics, TempO-Seq assays deliver an easy to use solution for customers doing expression profiling for any species.

TempO-Seq is unique in its capacity to avoid RNA purification or reverse transcription, by targeting RNAs with detector oligos and removing excess probes and enzymatic inhibitors before the first enzymatic step. Correctly hybridised detector oligos are ligated, then amplified through primer landing sites that are shared among all probes,

This approach permits high target multiplexing because although the central part of the ligated oligos contains diverse sequences, only two PCR primers are needed for any sample, eliminating the primer cross-hybridisation and competition inherent in multiplex PCR. Dual sequence tags are incorporated during PCR, to identify up to 6,144 samples in one sequencing library.