2026-10-11T13:57:37Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33960001 2026-09-21T21:06:36Z category_146 category_7 category_21 category_272 category_39 category_734 category_931 portal_63 item_type_2 month_year_09_2026

Robotic Centrifugal Microfluidic Automation Enables Library Preparation of Nine Samples with a Tenfold Reduction in Reagent Consumption for Decentralized Sequencing Trong Nguyen (12963380) Viola Dreyer (9693415) Yumi Kaku (25085596) Peter Juelg (11154372) Tobias Hutzenlaub (9361651) Stefan Niemann (62805) Nils Paust (6997982) Jacob Friedrich Hess (9361645) Space Science Medicine Biotechnology Environmental Sciences not elsewhere classified Ecology Biological Sciences not elsewhere classified Information Systems not elsewhere classified wet laboratory workflows substantially reduced due settings still rely placing required reagents employ standardized pipelines average coverage depth manual reference workflow costs per sample namely reagent preparation manual handling steps supports library generation mycobacterium tuberculosis </ lp platform combines involving complex steps existing automation approaches total ngs costs strains including drug decentralized sequencing next sample handling lp platform manual pipetting investment costs library retrieval generation sequencing extensively drug decentralized laboratories reagent consumption tenfold reduction tenfold decrease satisfying throughput samples investigated pipetting robot nine samples microbial genomes medical diagnostics mdr ), crucial tool critical bottleneck centrifugal microfluidic bcg vaccine based cartridge 98 %. 97 %, Next-generation sequencing (NGS) has become a crucial tool for medical diagnostics in decentralized laboratories. While sequencing and bioinformatics themselves employ standardized pipelines, wet laboratory workflows in these settings still rely on manual pipetting and sample handling. In particular, the library preparation procedures represent a critical bottleneck, accounting for up to 50% of total NGS costs and involving complex steps. Automation offers potential solutions to these challenges. However, existing automation approaches have limitations in satisfying throughput, investment costs, costs per sample, and degree of automation. Here, we implement a robotic centrifugal microfluidic platform around the Illumina Nextera XT DNA Library Preparation Kit or RoCM-LP platform, which supports library generation from microbial genomes. The RoCM-LP platform combines a centrifugal microfluidic-based cartridge with a pipetting robot, capable of producing nine libraries per run at low-medium initial investment costs ($50k–70k). The costs per sample were substantially reduced due to a tenfold decrease in reagent consumption in comparison to the manual reference workflow, and the number of manual handling steps was minimized from 45 to just three, namely reagent preparation (placing required reagents into the RoCM-LP platform), sample loading, and library retrieval. The approach achieved quality metrics comparable to those of the manual reference workflow and met all required sequencing quality thresholds across four <i>Mycobacterium tuberculosis</i> complex (MTBC) strains including drug-susceptible, BCG vaccine (Pasteur Bacillus Calmette–Guérin), multidrug-resistant (MDR), and extensively drug-resistant (XDR) strains for tuberculosis (TB) samples investigated, with an average coverage depth of 107, an average mapped read percentage of 97%, and an average coverage breadth percentage of 98%. 2026-09-21T00:00:00Z Dataset Media 10.1021/acs.analchem.6c03501.s003 https://figshare.com/articles/media/Robotic_Centrifugal_Microfluidic_Automation_Enables_Library_Preparation_of_Nine_Samples_with_a_Tenfold_Reduction_in_Reagent_Consumption_for_Decentralized_Sequencing/33960001 CC BY-NC 4.0