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 I
- 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 an easy to use and 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 for quality Control
The Universal DNA Spike has a synthetic sequence that is not present in any known living organism. This DNA sequence is 1000 base pairs in length. 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 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 (Figure 2). The magnitude of the difference between these Cq values reflects the degree of inhibition.