Sequins built for RNA sequencing.

Synthetic RNA standards representing spliced gene isoforms.

Synthetic genes.

RNA sequencing can measure gene expression and assemble spliced isoforms. However, transcriptome complexity, expression-dependent biases, and other technical variables confound the analysis of RNA sequencing.

To address these challenges, we have built a set of synthetic splided genes, termed sequins, that act as internal controls during RNA sequencing.

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Using sequins with RNA sequencing.

Sequins are ‘spiked-in’ to your RNA sample at a fractional concentration. The sample, with the sequins, then undergoes reverse-transcription, library preparation and sequencing together.

Because of their synthetic sequence, the sequins can be distinguished from the sample in the output library. The sequins can then be analyzed as internal controls for your RNA sequencing experiment.

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Sequin analysis.

RNA sequins are mixed at staggered concentrations to form a reference ladder that can be used to evaluate the sensitivity and quantitive accuracy of your RNAseq library.

Sequins can assess the impact of technical variables, benchmark bioinformatic tools, and improve the accurate  analysis of your RNA sample.

Further reading.

Scientific paper
Spliced synthetic genes as internal controls in RNA sequencing experiments. (2016)
Hardwick et. al.,
Nature Methods.

Protocol
Using sequins with RNA sequencing.
Laboratory protocol.
Version 2.1
October 2018