INTRODUCING TEMPO-SPAN
Key features
TempO-SPan (Templated Oligo-Sequencing Pandemic) is based on hybridization and sequencing of highly specific detector oligos.
input down to 10 pg with single-base specificity
up to 20 targets
no RNA extraction required, cDNA conversion or preamplification
no proprietary instrumentation: standard PCR cycler or microplate incubator is all you need
group up to 96 samples in one well and uniquely identify positive samples
maximize sequence capacity due to short barcode reads: up to 6,144 samples in one run
No change to “LIMITATIONS OF RNA-SEQ” OR “DETECTOR OLIGO DESIGN”
SARS-CoV-2, Influenza A/B, and RSV available now
TEMPO-SPAN WORKFLOW
The workflow for TempO-SPan is very similar to TempO-Seq™ with the exception that TempO-SPan adds an additional layer of barcoding to ensure that each sample is uniquely identifiable. With TempO-Seq, identifying the thousands of genes and the number present is the objective, with TempO-SPan, identifying if thousands of samples have a particular targeted gene is the objective.
Based on BioSpyder Technologies’ proprietary Templated Oligo Detection Assay, TempO-SPan 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.
For clinical evaluation, we have developed a SARS-CoV-2 panel that shows high sensitivity and specificity, while maintaining ease of use and reducing labor and consumables costs. The assay can detect fewer than 10 copies of viral RNA, equivalent or better than low throughput real time quantitative PCR tests.
TempO-SPan is customizable in the same way as TempO-Seq, by designing detector oligos for the desired content. A respiratory panel that includes SARS-CoV-2, Influenza A/B, and RSV is also available.
TempO-SPan has all the other benefits of TempO-Seq, including the ability to use crude sample lysates and simplified data alignment and analysis – see TempO-SeqR. For application to clinical testing, we have developed automated calling software to generate patient reports.
Like TemO-Seq, TempO-SPan 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 hybridized detector oligos are ligated, then amplified through primer landing sites that are shared among all probes.
