Automated HMW-DNA extraction validated on Tecan robot

Further optimisation of the multi-well filter plates is ongoing in RevoluGen’s R&D laboratories with commercial launch of the automated Fire Monkey HMW-DNA extraction kit anticipated in the coming months

RevoluGen has announced the technology validation of the automated extraction of library-ready high molecular weight (HMW) DNA in a multi-well filter plate format.

Under a collaboration agreement with Tecan, announced in 2020, RevoluGen has completed the configuration and validation testing (by DNA sequencing) of an automated workflow using an adapted Tecan Fluent 780 robotic liquid handler with an integrated Rotanda 460 robotic centrifuge using laboratory standard multi-well plates, with patented Fire Monkey chemistry and matrix, enabling automation of the spin-based process.

The automated HMW-DNA extraction protocol has been validated to extract and purify bacterial samples. Further optimisation of the multi-well filter plates is ongoing in RevoluGen’s R&D laboratories with commercial launch of the automated Fire Monkey HMW-DNA extraction kit anticipated in the coming months.

“RevoluGen's automation of DNA extraction, the critical first step in the DNA sequencing process, comes at a good time as structural variant surveillance could drive a sequencing boom” said Revolugen’s CEO, Pieter Haitsma Mulier.

In the validation run to demonstrate quality of sequencing output, eight E. coli cell suspensions were transferred to a 96-well plate and, after a fully hand-free process, the output DNA delivered sequencing results equivalent to manually-extracted Fire Monkey spin-column based HMW-DNA. The average strand length achieved was 106kb with sequencing on the Oxford Nanopore (ONT) MinION LSK110 platform delivering more than 30Gb at N50 longer than 30kb, with good pore health.

The nucleic acid isolation and purification (NAIP) technology is based on a spin-based protocol to extract HMW-DNA using a high g-force that does not break the long and fragile DNA molecules. It produces DNA containing fragments that are not too short or long, the company claims, improving the overall sequencing process by not wasting resources.

“Extracting HMW-DNA based on a laboratory standard spin-column process was revolutionary in itself. Given the demand for automation, we have worked with Tecan to achieve a working robotic system that fully automates the process for DNA extraction of long length DNA using an adapted multi-well plate process. This is a truly disruptive innovation and will support massively high throughput, high quality, cost effective DNA sequencing” said Dr Georgios Patsos, inventor of the Fire Monkey technology and CSO at RevoluGen.

The rapid growth of sequencing worldwide requires automated NAIP and sequencing processes to handle the volume of samples needed in applications such as population genomics, epidemiological mutation screening and antibiotic microbial resistance gene monitoring.

Dr Gemma Langridge, a Group Leader at the UK’s Quadram Institute, who has been an early adopter of RevoluGen’s Fire Monkey technology said: “With the automation that is coming from RevoluGen, what we are looking for is to be able to prep hundreds of bacterial strains at a time, then the scope for our research increases quite dramatically that way.”

Currently every sequencing run requires each DNA sample to be extracted individually. A validated and automated Fire Monkey process for multiplexed automated HMW-DNA extraction will further lower the overall cost of sequencing, Revolugen claims, and thus accelerate the uptake of sequencing in many high-volume dependent applications.

“Automation represents a game-changing differentiator and will allow us to rapidly scale sequencing at a lower cost per unit. Now that we have validated the automated protocol, we believe that Fire Monkey is poised to become the DNA extraction technology of choice for high volume applications. Our technology unites short-read and long-read sequencing, enabling both from the same sample. It provides flexibility to revisit the same original sample for more detailed long-read sequencing after a first run with cheaper short-read sequencing” concludes RevoluGen CEO, Pieter Haitsma Mulier.

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