•w
w
a
ta
ta
w.
.co
The inhibition of the qPCR reaction causes erroneous biological readouts, as the degree of inhibition varies between
samples. The results of qPCR are often at risk of being compromised by contaminants present in samples that can
inhibit enzymatic reactions and by degradation of DNA during sampling, transport and storage. The Universal DNA
Spike from TATAA Biocenter is an easy to use and very effective tool for quality control of your qPCR experimental
workflow.
Test for inhibition
Validation using the Universal DNA Spike is effective, easy to perform and provides an important additional
assurance that both the pre-analytical and analytical phases of your experiment are valid. The measured Cq-value
and the shape of the amplification curve reflect inhibition and/or losses during extraction, handling, transport and
storage of samples, including freeze-thaw events during qPCR. If the Cq-value is greater in an experimental sample
than in the control then you know that the analytical process of that experimental sample is inhibited (Figure 1.).
A.
B.
Figure 1. Plot A) shows a qPCR amplification plot with no inhibition as the Spike control in water and in sample matrix
shows the same Cq value and efficiency. In B) the qPCR inhibition is seen on the lower slope of the amplification curve.
Test for extraction yield
The TATAA Universal DNA spike may be added to any stabilised and homogenised sample prior to DNA purification
to test for material loss during isolation, transportation, and storage.
Equal quantities of the DNA spike are added to each experimental sample and to a control sample. The control
sample should be based on nuclease free water or elution buffer of the same volume as used for elution/dissolving
in DNA purification protocol. An equal Cq of the TATAA Universal Spike in the experimental and control samples
reflects 100% yield. If the Cq values differ, the yield of the extraction can be estimated using the following formula:
1
yield (%) = ____________________
100
(Cq experimental sample - Cq control) *
2
Ordering information
The TATAA Universal DNA Spike can be ordered from our distribution partners around the world or in our webshop.
Please visit www.tataa.com.
The TATAA Universal DNA Spike is available both as Probe and SYBR kits. A 6 months complimentary license for
GenEx Enterprise is included with the kit.
TATAA Biocenter AB
Odinsgatan 28, 411 03 Göteborg
Tel: +46 31 761 57 00, Fax: +46 31 15 28 90, E-mail: [email protected]
•
The risk of compromised qPCR results
m
TATAA Universal DNA Spike - SYBR/Probe
•w
w
a
ta
ta
w.
.co
m
TATAA Universal RNA Spike - SYBR/Probe
•
The risk of compromised RT-qPCR results
The inhibition of the RT-qPCR reaction causes erroneous biological readouts, even though the qPCR amplification
curves look normal. The results of RT-qPCR are often at risk of being compromised by contaminants present in
samples that can inhibit enzymatic reactions and by degradation of RNA and cDNA during sampling, transport
and storage. The Universal RNA Spike from TATAA Biocenter is an easy to use and very effective tool for quality
control throughout the entire RT-qPCR experimental workflow.
Test for inhibition
Validation using the Universal RNA Spike is effective, easy to perform, and provides an important additional
assurance that both the pre-analytical and analytical phases of your experiment are valid. The measured Cq and the
shape of the amplification curve reflect inhibition and/or losses during extraction, handling, transport and storage
of samples, including freeze-thaw events during RT-qPCR. If the Cq-value is greater in an experimental sample than
in the control then you know that the analytical process of that experimental sample is inhibited (Figure 1.). The
magnitude of the difference between these Cq values reflects the degree of inhibition.
A.
B.
C.
D.
Figure 1. Plot A) shows a qPCR amplification plot with no inhibition as the Spike control in water and in sample matrix
shows the same Cq value and efficiency. In B) the RT-reaction is inhibited but not the qPCR reaction, seen on the
higher Cq-value. In C) the RT-reaction step has not been inhibited but the qPCR reaction has been as seen by the lower
slope of the amplification curve. In D) both the RT-step and qPCR reaction are inhibited.
Test for extraction yield
The TATAA Universal RNA spike may be added to any stabilised and homogenised sample prior to RNA purification
to test for material loss during isolation, transportation and storage.
Equal quantities of the RNA spike are added to each experimental sample and to a control sample. The control
sample should be based on nuclease free water or elution buffer of the same volume as used for elution/dissolving in the RNA purification protocol. An equal Cq-value of the TATAA Universal RNA Spike in the experimental and
control samples reflects a 100% yield. If the Cq-values differ, the yield of the extraction can be estimated using the
following formula:
1
yield (%) = ____________________
100
(Cq experimental sample - Cq control) *
2
Ordering information
The TATAA Universal RNA Spike can be ordered from our distribution partners around the world or in our webshop.
Please visit www.tataa.com.
The TATAA Universal RNA Spike is available both as Probe and SYBR kits. A 6 months complimentary license for
GenEx Enterprise is included with the kit.
TATAA Biocenter AB
Odinsgatan 28, 411 03 Göteborg
Tel: +46 31 761 57 00, Fax: +46 31 15 28 90, E-mail: [email protected]