How do I select the right SIRV spike-in set?

The choice of the SIRV spike-in set mainly depends on the aim of the study and the type of RNA of interest.

The table below summarizes the compatibility and ideal use cases of the different SIRV spike-in sets.

Table 1 | SIRV-Set Selection Guide

SIRV-Set 1

This set contains three different mixes (E0, E1 and E2) each including the same 69 isoforms but at different concentration ranges. This set is ideal for fold-change measurements, isoform detection and identification, calibration of wet lab experiments and validation of data analysis pipelines.

SIRV-Set 2

This set contains SIRV mix E0 in which all 69 isoforms are present at equimolar concentrations. This set is ideal for isoform discovery and quantification with both short-read and long-read sequencing applications (effective even at low sequencing depth). This set is recommended if ERCCs are spiked-in separately at other steps in the workflow (e.g. prior to RNA extraction, or enrichment/deletion steps).

SIRV-Set 3

This set combines ERCC spike-ins with the SIRV isoforms (mix E0) creating a comprehensive all-purpose control set. It allows absolute quantification and cross-sample normalization providing concentration range measurements (ERCC) and isoform resolution (SIRVs).

SIRV-Set 4

This set includes ERCCs, SIRV isoforms (mix E0) and additional long SIRVs ranging from 4kb to 12kb making it suitable for long read-sequencing approaches.