The Spike-In RNA Variants (SIRVs) are sets of transcript designed to validate the performance of RNA sequencing workflows, especially how bioinformatics algorithms correct biases introduced by library preparation and sequencing. This SIRV-Set 1 contains three isoform mixes, E0, E1 and E2,which contain the same synthetic transcripts but in different concentration ratios. 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
- In-depth validation of isoform detection and quantification pipelines
- Validation of mRNA quantification workflows
- Assessment of differential gene expression on the isoform level
- The kit includes 1 tube each of the SIRV mixes E0, E1 and E2, which contain 69 SIRV isoform transcripts in 3 different concentration ranges.
- The 4 µl content of each mix is sufficient to use safely 3 times 1 µl for spiking in hundreds of, e.g., 100 ng total RNA-Seq samples (or millions of single-cell RNA-Seq samples).
- Molecular-biology grade water for dilution of the RNA.
- Compatible with almost all RNA-Seq protocols, including polyA-selective methods (only excluding Cap-specific preparations).