The PCR decontamination kit uses a dsDNase to remove contaminating DNA from mastermixes. The double-strand specific property allows decontamination in the presence of primers and probe.The dsDNase is irreversibly inactivated by heating to 60°C in the presence of DTT, ensuring that DNA template added to the mastermix after inactivation will not be digested. The mastermix can be used immediately after decontamination to run PCR reactions.
AMPIGENE® Taq polymerases have been treated to remove host DNA and additional clean-up is not necessary. The PCR decontamination kit is recommended for decontamination of primers or competitor’s PCR mixes.
Fig 3. Simple workflow for decontamination
Fig 2. Untreated and decontaminated qPCR 2x master mix was used for analysis of an E. coli gDNA 10-fold serial dilution with 5 steps. NTC samples were included, and all plots of the serial dilution show an average of three replicates. Inset: Standard curves calculated from Cq values obtained from analysis of serial dilution.
Fig 1. The presence of E. coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water as template (NTC) and following the manufacturer’s instructions. The figure shows plots acquired from several separate experiments. Traces of E. coli 23S DNA were found in all master mixes tested, with Cq values generally ranging from 30 – 35.
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Product Details
Application Notes:
For the removal of contaminating DNA from primers and mastermixes prior to PCR.
Shipping:
Blue Ice
Short Term Storage:
-20°C
Long Term Storage:
-20°C
Contents:
Heat-labile dsDNase DTT
Scientific Background:
Polymerases used in PCR are frequently contaminated with E. coli DNA. Contaminating DNA may cause reduced sensitivity and false positives when small amounts of bacterial DNA are targeted. Other sources of contamination might be dNTPs, buffer components and primers / probes, as well as DNA introduced during handling. The PCR decontamination kit is an easy method to eliminate DNA contamination.
Technical Info/Product Notes:
The PCR decontamination kit is not recommended for use with high fidelity PCR buffers. High fidelity PCR buffers typically contain high salt concentration, low magnesium concentration, and high pH which reduce the activity of dsDNase.
UniProt ID:
C9YSL6_PANBO
Regulatory Status:
RUO - Research Use Only
Product Literature References
Signatures of HIV and Major Depressive Disorder in the Plasma Microbiome: B.C. Taylor, et al.; Microorganisms 11, 11022 (2023), Abstract;
