a qPCR amplification plot with no inhibition as the Spike control in water and in sample matrix shows the same Cq value and efficiency
The qPCR inhibition is seen on the lower slope of the amplification curve. If the qPCR is inhibited, results are unreliable.
TATAA Universal DNA Spike 166 bp
- Reactions/SYBR or Probe:
Control for qPCR-inhibition
The Universal DNA Spike from TATAA Biocenter is easy to use and an very effective tool for quality control of the qPCR experimental workflow. DNA Spike is used in the qPCR reaction to detect any inhibition. In addition TATAA Universal RNA Spike I and II can be used to detect any inhibition during the cDNA synthesis reaction.
In cases where predominantly short fragments of DNA may be present such as in free circulating DNA or in heavily degraded material a shorter DNA spike template , like Spike 166 bp or Spike 500 bp, may more correctly resemble DNA present in samples.
The Universal Spikes from TATAA Biocenter are offered as RNA or DNA templates in different synthetic sequences that are not present in any known living organism and can be used as very effective tools for quality control throughout the entire RT-qPCR experimental workflow.
TATAA Universal DNA Spike 166 bp has a DNA sequence length of 166 base pairs. The Spike 166 bp assay which is used for detecting the Universal DNA Spike 166 bp template, is very robust and optimized for high sensitivity for inhibition. The assay amplifies a 69-base region of the synthetic template. The measured Cq-values and the shape of the amplification curves reflect the inhibition. The Cq-values of the Spike 166 bp assay also reflect losses during extraction, handling, transport, and storage of samples, including freeze-thaw events during RT-qPCR.