ERCC RNA Spike-In Mix - FAQs

View additional product information for ERCC RNA Spike-In Mix - FAQs (4456740)

14 product FAQs found

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.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Gene Expression Support Center.

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)