ATTENTION: All SIRV-Sets are currently shipped on dry ice and should be stored at, or below, -20°C upon arrival. No resolubilization is required. Please see User Guide for detailed information on changed shipment, storage, and usage conditions.
The Spike-In RNA Variants (SIRVs) are sets of transcript designed to validate and control the performance of RNA sequencing workflows, especially how bioinformatics algorithms correct biases introduced by library preparation and sequencing. This SIRV-Set 3 combines the equimolar SIRV isoform mix E0 and one ERCC mix. The E0 mix provides controls for resolving isoform complexity by containing 69 SIRV isoforms at the same molarity. This is complemented by 92 non-isoform ERCC transcripts of unique sequence identity at a concentration range of 6 orders of magnitude. The spike-ins can be used with different RNA inputs (total RNA, poly(A) selected RNA, or rRNA-depleted RNA), library preparation methods and sequencing platforms.
Features and Application:
- Comprehensively reflects variations of alternative splicing, alternative transcription start- and end-sites, overlapping genes, and antisense transcripts, as well as variations of concentrations
- Efficient evaluation of isoform-specific workflows
- Covers the entire dynamic range of endogenous RNA
- Applicable to pipelines with low read-depth
- Validation and control of mRNA quantification workflows probing isoform detection and concentration measures
- Evaluation of experiment concordance to assess the comparability of data sets
- The kit contains 1, or 3, tube(s) with SIRV isoform mix E0 combined with one ERCC mix.
- The dried content of each tube has to be reconstituted in water and is sufficient to spike at least 9 times dozens to hundreds of RNA-Seq samples (or millions of single-cell RNA-seq samples).
- Molecular-biology grade water for reconstitution and dilution of the RNA.
Compatible with almost all RNA-Seq protocols, including polyA-selective methods (only excluding Cap-specific preparations).