Ultra-sensitive detection of DNA or RNA from complex, inhibitor-rich blood samples
Sensitivity of DNA Amplification in the Presence of 20% Serum
Activity of lyophilized Lyo-Ready ™ Direct DNA qPCR Blood ( red ) was compared to KAPA Probe Force ( green ) in a multiplex qPCR assay, using a 10-fold serial dilution of plasmid DNA (10,000, 1000, 100 and 10 copies respectively), in the presence of 20% serum. The results illustrate that Lyo-Ready ™ Direct DNA qPCR Blood has significantly higher sensitivity and reproducibility than KAPA Probe Force in assays detecting high or low copies of DNA template and in the presence of PCR inhibitors.
10,000 copies 1,000 copies
100 copies 10 copies
Lyo-Ready ™ Direct DNA qPCR Blood | KAPA Probe Force
Sensitivity of RNA Amplification in the Presence of 5% K2-EDTA Whole Blood
Lyo-Ready ™ Direct RNA/DNA qPCR Blood ( red ) was compared to UltraPlex 1-Step ToughMix ( grey ) in a multiplex qPCR assay, using a 10-fold serial dilution of mammalian RNA (10,000, 1000, 100 and 10 copies respectively), in the presence of 5% K2-EDTA whole blood. The results illustrate that Lyo-Ready ™ Direct DNA qPCR Blood has significantly higher sensitivity and reproducibility than UltraPlex in assays detecting high or low copies of RNA template and in the presence of PCR inhibitors.
10,000 copies 1,000 copies
100 copies 10 copies
Lyo-Ready ™ Direct RNA/DNA qPCR Blood | UltraPlex 1-Step ToughMix
Significantly greater tolerance to inhibitors found in whole blood and anticoagulants (K2-EDTA, sodium heparin, and sodium citrate)
A/ Lyo-Ready™ Direct DNA qPCR Blood (MDX122)
45 B/ Lyo-Ready™ Direct RNA/DNA qPCR Blood (MDX123)
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25
2%
5%
10%
2%
5%
10%
2%
5%
10%
0%
2%
5%
10%
0%
2%
5%
10%
0%
2%
5%
10%
MDX122
KAPA Probe Force (Roche)
TaqPath ™ (ThermoFisher)
MDX123
UltraPlex ® (Quantabio)
TaqPath ™ (ThermoFisher)
EDTA | Heparin | Citrate
Anticoagulants and high concentrations of whole blood are known to inhibit qPCR efficiencies. In this experiment, 0% 2%, 5% and 10% whole human blood in the presence of K2-EDTA ( orange ), sodium heparin ( green ) or sodium citrate ( grey ) was tested with (A) Lyo-Ready ™ Direct DNA qPCR Blood (MDX122) against Kapa Probe Force (Roche) and TaqPath (Thermo) and (B) Lyo-Ready ™ Direct RNA/DNA qPCR Blood (MDX123) against UltraPlex 1-Step ToughMix (QuantaBio) and TaqPath 1-Step Multiplex Master Mix (Thermo). The results demonstrate that the reaction efficiencies of the Lyo-Ready Direct Blood mixes are higher in the presence of inhibitors found in anticoagulants and high concentrations of blood.
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