Ion AmpliSeq ERCC Companion Panel

The Ion AmpliSeq RNA ERCC Companion Panel works with the ERCC RNA Spike-In Mix to provide performance assessment of Ion AmpliSeq Transcriptome libraries in gene expression experiments. Add the ERCC RNA Spike-In Mix to each RNA sample, then amplify targets with the Ion AmpliSeq Transcriptome panel plus the Ion AmpliSeq RNA ERCC Companion Panel during library preparation. When analyzing the sequencing results, the data from sequenced ERCC targets is compared to the known concentration of the ERCC targets in the Spike-In Mix.

Working with the ERCC RNA Spike-In Mix, the Ion AmpliSeq RNA ERCC Companion Panel offers:
• Standard baseline of Ion AmpliSeq Transcriptome gene expression experiments across multiple experiments and various platforms
• Measurement of the lower limit of detection and dynamic range of an expression experiment
• Identification of degraded or low-yield samples by assessing the performance of ERCC targets

Variation in RNA expression data can be attributed to a variety of factors including the quality of the starting material, the level of cellularity and RNA yield, the platform employed, and the person performing the experiment. To control for these sources of variability, a common set of external RNA controls has been developed by the External RNA Controls Consortium (ERCC), an ad-hoc group of academic, private, and public organizations hosted by the National Institute of Standards and Technology (NIST). The controls consist of a set of unlabeled, polyadenylated transcripts designed to be added to an RNA analysis experiment after sample isolation, in order to measure against defined performance criteria.

The Ion AmpliSeq RNA ERCC Companion Panel consists of 10 primer pairs targeting 10 selected transcripts from the ERCC RNA Spike-In Mix representing a 13log₂ dynamic range. Chosen for general GC balance and amenability for linear amplification (TaqMan qPCR), the selected ERCC targets produce R² >0.9 under normal experimental conditions.