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Invitrogen™ ERCC RNA Spike-In Mix

Catalog No. 4456740
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Catalog No. 4456740 Supplier Invitrogen™ Supplier No. 4456740
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ERCC Spike-In Control Mixes are commercially available, pre-formulated blends of 92 transcripts, derived and traceable from NIST-certified DNA plasmids. The transcripts are designed to be 250 to 2,000 nt in length, which mimic natural eukaryotic mRNAs.

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. Up until the design of such universally accepted controls, it has been difficult to execute a thorough investigation of fundamental analytical performance metrics. From the trusted brand of quality RNA reagents, Ambion™ ERCC Spike-In Control Mixes are commercially available, pre-formulated blends of 92 transcripts, derived and traceable from NIST-certified DNA plasmids. The transcripts are designed to be 250 to 2,000 nt in length, which mimic natural eukaryotic mRNAs.

Key product features:

  • Achieve a standard measure for data comparison across gene expression experiments
  • Measure sensitivity (lower limit of detection) and dynamic range of an experiment
  • Quantitate differential gene expression

Unlock the Potential of RNA Analysis
RNA analysis, including gene expression profiling and whole transcriptome surveying, can lead to better understanding of expression patterns in disease states and provides greater insights into biological pathways and molecular mechanisms that regulate cell fate, development, and disease progression. Traditional methods of RNA analysis, such as qRT-PCR and microarrays, are well established but are being replaced by next-generation sequencing, a high-throughput digital alternative. Because each method carries multiple platforms, and with the need to compare various samples across platforms throughout the world, a standard measure for data comparison is necessary. As the capabilities of RNA analysis expand, the necessity to create a standardized view of data will become even more important.

Achieve and Compare Results with Confirmed Accuracy
Ambion™ ERCC RNA Spike-In Controls are used to create a standard baseline measurement of RNA both within an experiment and across multiple experiments performed using various samples and platforms. With two spike-in mix formulations, various measurements, such as sensitivity or dynamic range, can be examined to assess different parameters in an experiment or across experiments. Furthermore, expression fold-change ratios between two samples can be calculated with a high degree of confidence using the highly concordant relationship between ExFold RNA Spike-In 1 and ExFold RNA Spike-In 2. Measurements are determined via known molar concentrations for each transcript within a spike-in mix and through association of the two mixes using a combination of ratios across 4 different subgroups of the 92 transcripts. The controls are ideal for next generation sequencing experiments, such as on SOLiD™ System, and supported microarray platforms such as the Illumina™ Sentrix™ BeadChip.

Choose Among Flexible Options for ERCC Kit Configurations
Whether measuring dynamic range or gene expression fold-change, Ambion™ ERCC Spike-In Control Mixes are available in two kit configurations to meet your experimental needs. Use the ERCC Spike-In Mix to determine the dynamic range and lower limit of detection on your platform, and use the ERCC ExFold Spike-In Mixes to assess the accuracy of differential gene expression measurements.

Specifications

Content And Storage This kit contains:
• 10 μl ERCC RNA Spike-In Mix 1
• 1.75ml Nuclease-free Water

-Do not store the Spike-In Mixes in a frost-free freezer.

-Store ERCC RNA Spike-In Mix 1 at –20°C
-Store Nuclease-free Water at any temperature
Format Tube
Control Type RNA Control
For Use With (Application) Next-Generation Sequencing
Includes Control Template(s)
Product Line Ambion
Product Type ERCC RNA Spike-In Mix
Quantity 1 Kit
How do I analyze data from the ERCC controls?

There is a specific ERCC_Analysis Plugin for Torrent Suite Software. Please see the ERCC_Analysis Plugin User Bulletin (Publ. No. 4479068) for detailed information.

Where can I find the ERCC concentrations?

Please go to the ERCC RNA Spike-In Mix (Cat. No. 4456740) or ERCC Ex-Fold Spike-In Mix (Cat. No. 4456739) product page on www.thermofisher.com. Downloadable files are under the manuals section heading, click on Manual: ERCC Controls Analysis: ERCC RNA Spike-In Control Mixes.

Is there an ERCC-like product for bacteria or small RNA (or other non-polyA RNA species)?

No.

What is the difference between the ERCC RNA Spike-In Mix (Cat. No. 4456740) and the ERCC Ex-Fold Spike-In Mix (Cat. No. 4456739)?

The ERCC RNA Spike-In Mix (Cat. No. 4456740) includes Spike-In Mix 1 alone; it can be used to assess platform dynamic range and lower limit of detection. The ERCC Ex-Fold Spike-In Mix (Cat. No. 4456739). includes both Spike-In Mix 1 and Spike-In Mix 2. In addition to the performance measurements described above for the ERCC RNA Spike-In Mix, this kit can be used to assess the accuracy of measurements of differential gene expression on your platform. Please see the ERCC RNA Spike-In Control Mixes User Guide (Pub. No. 4455352) for detailed information.

What are the ERCC RNA Spike-In Control mixes (Cat. No. 4456740 & 4456739)?

The ERRC RNA Spike-In Control mixes are a set of external RNA controls developed by the External RNA Controls Consortium (ERCC) and NIST (National Institute of Standards and Technology). The set is comprised of 92 synthetic, unlabeled polyadenylated transcripts of various lengths for determining the limit of detection (LOD) and the dynamic range for mRNA expression studies (in eukaryotes). The spike-in mix should be added to the RNA sample after isolation but before making the library. For more information, please refer to ERCC RNA Spike-In Control Mixes User Guide (Publ. No. 4455352) and ERCC_Analysis Plugin User Bulletin (Publ. No. 4479068).

Does the ERCC RNA Spike-In Mix (Cat. No. 4456740) contain only single-stranded RNA or is there also double-stranded RNA?

The ERCC RNA Spike-In Mix (Cat. No. 4456740) contains only single-stranded RNA.

Does ERCC (External RNA Controls Consortium) RNA Spike-In Mix (Cat. No. 4456740) use a positive or negative strand control?

ERCC (External RNA Controls Consortium) RNA Spike-In Mix (Cat. No. 4456740) only contains positive-strand transcripts.

How many reactions can I perform with the Ion AmpliSeq RNA ERCC Companion Panel?

The Ion AmpliSeq RNA ERCC Companion Panel provides enough reagents to perform 96 manual reactions or 48 automated reactions.

What software do I need to view data from the Ion AmpliSeq RNA ERCC Companion Panel?

To view data from the Ion AmpliSeq RNA ERCC Companion Panel, Torrent Suite Software version 5.10 or later is required and Version 5.10.0.3 or later of the ERCC_Analysis plugin for Torrent Suite Software is required.

Can the Ion AmpliSeq RNA ERCC Companion Panel be used with FFPE samples?

We do not recommend using the Ion AmpliSeq RNA ERCC Companion Panel with FFPE samples or with any panels besides human and mouse Ion AmpliSeq transcriptome gene expression panels.

How much of the Ion Ampliseq RNA ERCC Companion Panel should I add to the target amplification reactions?

We recommend adding 1 µL of the Ion Ampliseq RNA ERCC Companion Panel per manual target amplification reaction and 8 µL per primer pool for an automated target amplification reaction on the Ion Chef Instrument.

Why would one use the Ion AmpliSeq ERCC Companion Panel?

The Ion AmpliSeq ERCC Companion Panel is compatible with Ion AmpliSeq Transcriptome Human Gene Expression Kit and Ion AmpliSeq Transcriptome Mouse Gene Expression Kit (not recommended for use with FFPE samples or panels outside of Ion AmpliSeq Human or Mouse Transcriptome kits). With the addition of the Ion AmpliSeq RNA ERCC Companion panel to your Ion AmpliSeq RNA panel, dynamic range and sensitivity can be similarly evaluated in Ion AmpliSeq RNA libraries that are prepared from RNA samples containing the ERCC RNA Spike-In Mix

How many primer pairs are in the Ion AmpliSeq RNA ERCC Companion Panel?

The Ion AmpliSeq RNA ERCC Companion Panel consists of 10 primer pairs targeting 10 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.

Which kits are compatible with the Ion AmpliSeq RNA ERCC Companion Panel?

The Ion AmpliSeq RNA ERCC Companion Panel is compatible with both manual and chef-ready versions of human and mouse Ion AmpliSeq transcriptome gene expression kits:

- Ion AmpliSeq Transcriptome Human Gene Expression Kit (Cat. No. A26325, A26326, or A26327)
- Ion AmpliSeq Transcriptome Mouse Gene Expression Kit (Cat. No. A36553, A36554, or A36555)
- Ion AmpliSeq Transcriptome Human Gene Expression Panel, Chef-Ready Kit (Cat. No. A31446)
- Ion AmpliSeq Transcriptome Mouse Gene Expression Panel, Chef-Ready Kit (Cat. No. A36412)


For Research Use Only. Not for use in diagnostic procedures.