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 II
- Reactions/SYBR or Probe:
Control for Inhibition of the qPCR - the risk of compromised RT-qPCR results
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.
TATAA Universal DNA Spike II which has a DNA sequence length of 2000 base pairs. The Spike II assay which is used for detecting the Universal DNA Spike II template, is very robust and optimized for high sensitivity for inhibition. The assay amplifies a 286-base region towards the 3’ end of the synthetic template. The measured Cq-values and the shape of the amplification curves reflect the inhibition. The Cq-values of the Spike II assay also reflect losses during extraction, handling, transport, and storage of samples, including freeze-thaw events during RT-qPCR.