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Article: Decontamination of PCR master mixes can improve accuracy and sensitivity in microbiome analyses

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

Article published in International Journal of Molecular Sciences

The team of Anders Ståhlberg (Sahlgrenska Cancer Center at University of Gothenburg) in collaboration with ArcticZymes has today published an article in International Journal of Molecular Sciences.

The article, titled “Preamplification with dUTP and Cod UNG Enables Elimination of Contaminating Amplicons”, describes how the use of dUTPs in combination with Cod UNG in targeted preamplification can provide a simple and efficient solution for eliminating carry-over contamination.

Carry-over contamination can pose significant challenges when analysing small samples of DNA and RNA where preamplification is required. By introducing contamination clean-up with Cod UNG prior to preamplification with dUTP in the work-flow, these challenges can be minimized.

The full article is available here.

ArcticZymes secures first supply agreement for utility of Salt Active Nuclease in Gene Therapy

ArcticZymes has secured its first supply agreement for the utility of Salt Active Nuclease High Quality (SAN HQ) in the cGMP manufacturing of viruses for gene therapy. ArcticZymes’ Novel SAN HQ enzyme offers gene therapy and vaccine customers a more cost effective and technically superior solution for removing contaminating DNA during the manufacturing process of therapeutic viruses. This represents a critical and challenging step in the manufacturing process where long-established alternative technologies are costlier and technically suboptimal. Cost and technical inefficiencies represent key challenges for the wider adoption of gene therapy. ArcticZymes SAN HQ allows customers to overcome these hurdles in one part of the complicated workflow in the manufacturing of cGMP grade therapeutic viruses. cGMP (current Good Manufacturing Practices) is relevant when a drug leaves the laboratory and enters into production, where scale, consistent quality and robustness are key requirements.

The undisclosed customer is a leading pioneer in the rapidly growing gene therapy market. Unlike traditional therapeutics, gene therapy provides realistic opportunity to cure devastating childhood genetic diseases and cancers by using viruses to modify DNA within a patient’s cells.

Commercial value of individual deals is anticipated to be in the 1-3 MNOK range per annum as customers lock down their large-scale cGMP manufacturing processes. The ability to produce viruses in large-scale is a critical factor to ensure that enough virus can be produced to support clinical trials and approved therapeutics.

“Securing our first supply agreement to support cGMP manufacturing of gene therapy viruses via our novel technologies is a milestone for ArcticZymes. We are excited and privileged that ArcticZymes can play a part in supporting our customers who are pioneering gene therapy solutions to cure devastating and life -threatening diseases.” says Jethro Holter, Managing Director ArcticZymes, AS.

ArcticZymes expands SAP and Cod UNG portfolio

ArcticZymes presents rSAP Glycerol-FREE and Cod UNG Triton-FREE. rSAP Glycerol-FREE becomes the fourth member of ArcticZymes’ glycerol-free product family, whereas the launch of Cod UNG Triton-FREE marks the first step towards maintaining a completely EU REACH compliant product portfolio after Triton X-100 becomes subject to authorisation in January 2021.

Glycerol-free formulations enable efficient lyophilization. Lyophilization of enzyme products helps overcome logistical challenges such as cold-chain distribution and storage, and limited product shelf-lives. Glycerol-free formulations are also preferable in highly automated processes, where the viscosity of high-glycerol formulations may pose challenges.

Triton X-100, a commonly used detergent across the life science industries, was included in Annex XIV of the EU REACH regulation in June 2017, with a “sunset date” set to January 4th 2021. Companies in the EU/EEC that wish to continue using Triton X-100 after the sunset date, will have to apply for authorisation unless the particular use is exempt. By launching Cod UNG Triton-FREE, ArcticZymes demonstrates its commitment to maintaining a REACH compliant product portfolio, and facilitates a smooth transition for our existing Cod UNG customers.