Spike-In RNA Variant Control Mixes (SIRV)

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Spike-In RNA Variant Control Mixes (SIRV)

RNA sequencing is used in many research projects as a powerful tool for transcriptome analysis and holds promise for expanding into diagnostic and therapeutic applications. To be analytically valid, a laboratory test must deliver accurate information in a reproducible and robust manner. Markers and controls are the only way to unambiguously determine the reliability of your experiment, and Lexogen Spike-In RNA Variants (SIRVs) enable the evaluation of both RNA-Seq experiment quality and comparability.

o    Universal markers for comparing and monitoring RNA-seq experiments
o    Validate RNA-seq workflows, identify error sources or biases and improve RNA-seq workflows from Library prep to data analysis
o    Compatible with all platforms and with RNA from any organism, down to single cell level

Spike-in Transcripts in RNA-seq

The proliferation of different RNA-seq platforms and protocols as well as the continuous efforts to translate NGS into clinical diagnostics has created the need for multi-functional spike-in controls. These are integrated and processed with real samples to enable monitoring and comparing key performance parameters (eg. sensitivity, input-output correlation, detection and quantification of transcript variants). The external controls are RNA molecules of known sequence that are added in pre-determined amounts to a sample. They are subjected to the same protocol steps and the endogenous RNA to be separated only at the final step of the data analysis.

Figure 1 ǀ Workflow for using spike-in controls in RNA-Seq. Spike-In RNA Variants (SIRVs) are defined synthetic RNA molecules that mimic the main aspects of transcriptome complexity. They are added in minuscule amounts to samples before library preparation to undergo the very same processing steps as the endogenous RNA. After mapping the reads to the combined artificial genome, the spike-in data are used to analyse quality metrics and to categorize the experiments. The dotted lines show the decision-making processes of deciding i) if the complete data set is worthy of further processing (or if an experiment needs to be repeated), and ii) which data sets have concordance that will permit meaningful comparison of the full data sets to each other.

The SIRV Isoform and ERCC Molecules

Transcriptomes are complex and comprise several RNA classes with their own properties. Spike-in controls must reflect these to be representative for a given experimental design. The SIRV’s were developed as a family of modules that offer tailored solutions for the control of RNA measurements. The SIRV Isoform module is available on its own (SIRV-Set 1 and SIRV-Set 2) as well as in combination with the ERCC Module (SIRV-Set 3).

SIRV Set Selection Guide

The Spike-In RNA Variants of the isoform and ERCC modules (see Modular Design) are realized as defined mixes, and for some modules, different mixes are available.

Mixes, and combinations thereof, are available in the form of SIRV sets. Table 1 presents a SIRV set selection guide to help you finding the right set of modules and mixes for your application.

Table 1 ǀ SIRV set selection guide. SIRV-Set 1 (Cat. No 025.03) contains the isoform mixes E0, E1 and E2 of the isoform module, SIRV-Set 2 (Cat. No 050.01 and 050.03) provides the isoform Mix E0 only, whereas SIRV-Set 3 (Cat. No 051.01 and 051.03) has the SIRV Isoform Mix E0 in a mixture with the ERCCs.   Yes-icon: applicable, no: not applicable, and partly applicable (or parts of the sets applicable).

Ordering information:
Contact us for pricing and additional information  
 

Cat. n°

Description

Size

025.03

SIRV-Set 1 (Iso Mix E0, E1, E2); 1 vial of each

3 vials

050.01

SIRV-Set 2 (Iso Mix E0)

1 vial

050.03

SIRV-Set 2 (Iso Mix E0)

3 vials

051.01

SIRV-Set 3 (Iso Mix E0/ERCC)

1 vial

051.03

SIRV-Set 3 (Iso Mix E0/ERCC)

3 vials

 

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