Description
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 4 combines the equimolar SIRV isoform mix E0, the ERCC mix, and the long SIRVs. 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 long SIRV module adds the length aspect to this control mix, covering sizes of 4 kb to 12 kb. 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
- Assess representation of transcripts > 2.5 kb in your RNA-Seq data
- Test the limit of RNA-Seq workflows with the long SIRV module in SIRV-Set 4 extending to 12 kb
- Evaluation of experiment concordance to assess the comparability of data sets
Kit Contents:
- The kit contains 1, or 3, tube(s) (Cat. No 141.01 and 141.03 respectively) with SIRV isoform mix E0 combined with ERCC mix and the long SIRVs.
- Molecular-biology grade water for reconstitution and dilution of the RNA.
System Compatibility:
- Compatible with almost all RNA-Seq protocols, including polyA-selective methods (only excluding Cap-specific preparations).