A paper recently published in Letters in Applied Microbiology describes advantages of using ArcticZymes’ PCR Decontamination Kit in master mix clean-up prior to PCR in microbiome analysis applications.
In molecular diagnostics and applied sciences, a current trend is to migrate from searching for a set of predefined microorganisms to identifying and quantifying all microorganisms present in the sample, known as the microbiome. Analysis of microbiomes are very sensitive to DNA contamination. PCR master mixes are commonly contaminated with small amounts of DNA which can confound sequencing data, particularly in cases where the sample is low in DNA.
The authors of the paper found that the main contributor to DNA contamination in their workflow was the master mix. By treating the master mix with ArcticZymes’ PCR Decontamination Kit prior to amplification, a 99% reduction in total number of contaminating sequences was observed. The authors conclude that decontamination of PCR reagents in microbiome analysis workflows could have a significant impact on both the accuracy and sensitivity when working with low-biomass samples.
ArcticZymes’ PCR Decontamination Kit contains dsDNase, a double-strand specific endonuclease that can be irreversibly inactivated by a short incubation step at moderate temperatures. Implementation of the PCR Decontamination protocol in sequencing workflows is easy, fast, and does not compromise the integrity of master mix-components such as polymerases or primers.
The PCR Decontamination is available in the webshop.
Stinson, L. , Keelan, J. and Payne, M. (2018), Identification and removal of contaminating microbial DNA from PCR reagents: impact on low‐biomass microbiome analyses. Lett Appl Microbiol. doi:10.1111/lam.13091