A variety of factors include the quality of the starting material, the level of cell structure and RNA yield, the platform used, and the people conducting the experiment. People, all contribute to the variability of RNA expression data. In order to control these variability factors, the External RNA Controls Consortium (ERCC), established by the National Institute of Standards and Technology, convened by academic, private and public organizations, developed a set of universal external RNA controls. Controls consist of a set of unlabeled polyadenylated transcripts that are designed to be added to RNA analysis experiments after sample isolation for comparison with set functional standards. Prior to the development of such widely accepted standards, thorough analysis of important analytical parameters was difficult. Ambion™ ERCC Internal Control Mix, from a trusted brand of high-quality RNA reagents, is a commercially available, pre-prepared mix of traceable 92 transcripts derived from NIST-certified DNA plasmids. The transcript is designed to be 250 to 2,000 nt in length, mimicking natural eukaryotic mRNA.

Main product features:

Standard determination to achieve data comparison between different gene expression experiments
• Determine the sensitivity (lower detection limit) and dynamic range of the experiment< br>• Quantifying Differential Gene Expression

Unlocking the Potential of RNA Profiling
RNA profiling including gene expression profiling and whole-transcriptome surveys can provide a better understanding of disease states. expression patterns and provide a deeper understanding of the biological pathways and molecular mechanisms that regulate cellular outcome, development, and disease progression. Traditional RNA analysis methods such as qRT-PCR and gene chip technology are already well established, but are gradually being replaced by high-throughput digital alternatives, namely next-generation sequencing methods. Because each method involves multiple platforms and requires comparison of a variety of samples across platforms from around the world, a standard means of data comparison is needed. As RNA analysis capabilities expand, the need to create standardized views of the data has become even more important.

Obtain and compare results with proven accuracy
Ambion™ ERCC RNA internal controls are used to neutralize in an experiment using multiple samples and platforms Create a standard baseline measurement of RNA between multiple experiments. Using two internal reference mixture formulations (Figure 1), multiple metrics such as sensitivity or dynamic range can be examined to evaluate different parameters within an experiment or between experiments (Figure 2). Moreover, the ratio of expression level amplitude changes in the two samples can be calculated with high confidence from the highly consistent relationship between ExFold RNA Spike-In 1 and ExFold RNA Spike-In 2 (Figure 3). Since the molar concentrations of the various transcripts in the internal control mixture are known, the measurement can be determined by correlating the two mixtures using different ratios of the 4 subpopulations of the 92 transcripts. These controls are ideal for next-generation sequencing experiments such as the SOLiD™ System and supported microarray platforms such as the Illumina™ Sentrix™ BeadChip.

ERCC kit configuration options are flexible
Whether measuring dynamic range or gene expression fold changes, Ambion™ ERCC Internal Reference Mix is available in two kit configurations to meet your experimental needs . Use the ERCC Spike-In Mix to determine the dynamic detection range and detection limit of your current platform, and use the ERCC ExFold Spike-In Mix to evaluate the accuracy of differential gene expression assay results. < tr>< td>1.75 mL

	ERCC RNA Internal Reference Mix 1*	ExFold Internal Reference Mix 1*	ExFold internal reference mixture 2*	Nuclease-free water
ERCC RNA Internal Reference Mix (Cat. 4456740)	10 µL	—	—
ERCC ExFold RNA Internal Reference Mix (Item 4456739)	—	10 µL	10 µL	1.75 mL

&ast; Although ERCC RNA Internal Reference Mix 1 and ExFold Internal Reference Mix 1 The included ERCC transcript formulation is the same, but ERCC RNA Internal Control Mix 1 cannot replace ExFold Internal Control Mix 1 for fold change assessment. Only use ExFold Internal Reference Mix 1 and Mix 2 from the same production batch number.

Research use only. Not intended for diagnostic use.