ERCC ExFold RNA Control Mix provides a set of external RNA controls to evaluate the performance of a variety of technology platforms used for gene expression experiments Evaluate. Add an internal control mix to each RNA sample and run samples containing the internal control mix on the platform. Internal control mixture data were then compared to known internal control mixture concentrations and ratios to evaluate dynamic range, lower limit of detection, and fold-change response of the platform. Many gene expression technologies can benefit from the use of ERCC ExFold RNA Reference Mix, including next generation sequencing (NGS), microarrays, and PCR-based assays. Microarrays must contain probes for interrogation of ERCC transcripts to benefit from the use of the ERCC ExFold RNA Internal Control Mix. Consult the array manufacturer for details. Main product features:

•Standard determination to achieve data comparison between different gene expression experiments
•Measure the sensitivity (lower limit of detection) and dynamic range of the experiment
•Quantify differential gene expression

Reasons for needing external RNA controls
A variety of factors, including the quality of the starting material, cell structure and level of RNA yield, the platform used, and the person performing the experiment, can cause RNA expression to vary. Differences in 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. ERCC ExFold 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.

Unlocking the Potential of RNA Profiling
RNA profiling, including gene expression profiling and whole-transcriptome surveys, can provide a better understanding of expression patterns in disease states and provide insights into regulatory cells Provide deeper understanding of biological pathways and molecular mechanisms underlying 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
ERCC ExFold RNA Reference Mix is used to compare multiple samples in one experiment using multiple samples and platforms. Create a standard baseline measurement of RNA between experiments. Using two internal reference mixture formulations (see figure), different metrics such as sensitivity or dynamic range can be examined to evaluate different parameters within an experiment or between experiments (see figure). Furthermore, the ratio of expression fold changes between the two samples can be calculated with a high degree of confidence from the highly consistent relationship between ExFold RNA Control 1 and ExFold RNA Control 2 (shown in the figure). 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, ERCC internal control mixtures are 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.

	ERCC RNA Internal Reference Mix 1*	ExFold Internal Reference Mix 1*	ExFold internal reference mixture 2*	Nuclease-free water
ERCC RNA internal control mix (item 4456740)	10 µL	—< /td>	—	1.75 mL
ERCC ExFold RNA Internal Reference Mixture (Product 4456739)	—	10 µL	10 µL	1.75 mL

* Although ERCC RNA Mix 1 and ExFold Mix 1 contain the same ERCC transcript formulation, ERCC RNA Mix 1 cannot replace ExFold Mix 1 for fold change assessments. Only use ExFold Internal Reference Mix 1 and Mix 2 from the same production batch number.