TATAA Universal RNA Spike I – Synthetic RNA Spike-In for Extraction and RT Efficiency Control

The TATAA Universal RNA Spike I is a synthetic RNA fragment designed to act as a consistent internal control in RNA workflows. Spiked into samples before RNA extraction or RT, it enables scientists to:

•     Evaluate RNA extraction efficiency
•     Monitor RT performance and enzyme inhibition
•     Control for sample-to-sample and inter-assay variation
•     Improve the reliability of gene expression results

It is ideal for low-input or degraded samples, including FFPE tissue and biofluids.

TATAA Universal RNA Spike for quality control

The Universal RNA Spike from TATAA Biocenter is easy to use and very effective tool for quality control throughout entire RT-qPCR experimental workflow. The Universal RNA Spike has a synthetic sequence that is not present in any known living organism. This RNA sequence is 1000 bases in length, which includes a 5’ Cap and a poly-A tail of approximately 200 bases, hence the TATAA Universal RNA Spike mimics eukaryotic mRNA.

The Spike assay, which is very robust and is optimized for high sensitivity for inhibition, amplifies a 300-base region towards the 3’ end of the synthetic transcript. The risk of compromised RT-qPCR results Contaminants present in samples are known to inhibit enzymatic reactions and, in the context of a reverse-transcription quantitative Polymerase Chain Reaction (RT-qPCR) assay, are fully capable of distorting reported measurements.

Such enzymatic reactions are a necessary part of the sample-preparation prior to the qPCR, such as nuclease/proteinase treatment and subsequent reverse transcription of mRNA to cDNA. Inhibition of the RT-qPCR often causes erroneous biological readouts, even though the qPCR amplification curves can look perfectly normal. The reason is that these upstream reactions usually are exposed to a higher concentration of inhibitors than the PCR itself.

The measured Cq and the shape of the amplification curve reflect inhibition. If the Cq value is greater in an experimental sample than in the control then the analytical process of that experimental sample is inhibited. The magnitude of the difference between these Cq values reflects the degree of inhibition.

Test extraction yield

The TATAA Universal RNA spike may be added to any stabilised and homogenized sample prior to RNA purification to test for material loss during isolation, transportation and storage of samples, including freeze-thaw events during RT-qPCR.

Equal quantities of the RNA spike are added to each experimental sample and to a control sample. The control sample should be based on RNase free water or elution buffer of the same volume as used for elution/dissolving in RNA purification protocol. An equal Cq of the TATAA Universal RNA Spike in the experimental and control samples reflects 100% yield. If the Cq values differ, the yield of the extraction can be estimated.

Overview

•     Validated synthetic RNA spike-in
•     Enables normalization across runs and conditions
•     Non-homologous to endogenous RNA
•     Trusted by researchers in regulated and high-sensitivity environments
•     Developed and quality-tested by TATAA Biocenter

Applications

•     Spike-in control for RNA extraction and RT
•     RT-qPCR and RT-dPCR assay validation
•     Sample prep standardization across labs
•     Compatible with tissue, cell lines, and liquid biopsies

User Manual

All RNA spikes from TATAA Biocenter