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

Kit Contents:

  • 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.

System Compatibility:

  • Compatible with almost all RNA-Seq protocols, including polyA-selective methods (only excluding Cap-specific preparations).