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firefly technical note: KAPA EvoPlus Automated Library Preparation

This technical note provides supporting information for automating the KAPA EvoPlus DNA Library Prep Kit on SPT Labtech's firefly liquid handler. These protocols are available to download from the firefly community. Here we provide information on protocol run times, consumables usage and present data generated using these protocols.

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Technical note
Fully automated bead-based High Molecular Weight DNA extraction for long-read sequencing

The growing popularity of long read sequencing (LRS) is driving up demand for High Molecular Weight (HMW) DNA. However, the extraction of HMW DNA has proven to be a barrier for routine uses. Many conventional methods are expensive, resource- and time-intensive, and do not produce DNA of sufficient quantity and quality for LRS. Moreover, they might be challenging to scale up due to not being automation-friendly or requiring specialized equipment and labware, like vacuum manifolds or wide bore tips, driving the costs up.

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Application note
Enhancing Core Facility Operations: Automated Liquid Handling for High-Throughput, High-Quality NGS Library Generation

In this webinar, researchers from the Biopolymers Genomics Core Facility (BPF) at Harvard Medical School will discuss automating the versatile NEBNext UltraExpress DNA Library Prep Kit using SPT Labtech’s firefly liquid handling platform to generate high-quality libraries for next-generation sequencing from a broad input range of DNA. The accessible, streamlined NEBNext UltraExpress workflow in tandem with user-friendly firefly creates accessible means for generating NGS libraries for Illumina DNA sequencing. The speakers will explain how the ease of use of the firefly allowed the NEBNext UltraExpress kit to be easily written and optimized for automation and to facilitate a more productive sequencing service.

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Webinar
Accelerating Drug Discovery at Totus Medicines: DNA-Encoded Library Screening with firefly®

DNA-encoded libraries (DELs) have emerged as a transformative technology within drug discovery. The exploration of large libraries of small molecule ligands as a collective rather than an individual has springboarded DELs to the forefront of high throughput screening. Building upon traditional DEL methods, Totus Medicines has developed a platform capable of identifying covalent small molecule ligands via high-throughput cell-based DEL screening. This poster outlines how Totus Medicines is streamlining hit identification, optimization, and technology development, to accelerate the discovery of novel therapeutic candidates using SPT Labtech’s firefly automated liquid handling platform. We present the metrics that have been collected over one year of screening using Totus Medicine’s proprietary covalent DELs to run over 2000 unique selections against a large number of targets, spanning diverse therapeutic areas.

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Poster
dragonfly® discovery: Pioneering Sustainable Liquid Handling in the Modern Laboratory

In this poster, we highlight the significant role that dragonfly discovery plays in reducing the environmental impact of liquid handling workflows. In our first case study, dragonfly discovery demonstrates a reduction in plastic pipette tip usage by over 99% compared to traditional air displacement pipetting systems when performing a 3456-run enzymatic assay operated by 6-factor space-filling Design of Experiment (DoE). In Next-Generation Sequencing (NGS) library preparation workflows, such as the NEBNext® Ultra™ II FS DNA Library prep kit presented in our second case study, dragonfly discovery achieves a plastic reduction of over 50%.

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Poster
Towards high-throughput low-cost sustainable ecotoxicity testing

Traditional whole animal-based toxicity testing is expensive, time-consuming, and unethical. Furthermore, these tests do not capture the full range of biological impacts. There is growing interest in developing alternative methods (New Approach Methods, NAMs), that are less resource intensive (lower costs, quicker assays, fewer animals), while also providing more data on the mechanism of toxicity. The objective of this work was to optimize and establish a high throughput in vitro transcriptomic-based toxicity testing platform for chemical and environmental risk assessment. The platform couples human Caco-2 and Hep G2 cell lines with QIAGEN’s UPXome ultraplex technology, so that the combined test system can also derive transcriptomic points of departure (tPODs).

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Poster
Auto-tRNAseq: Driving throughput in tRNA library generation with firefly®

Transfer RNAs (tRNAs) are the critical adaptor molecules that translate genetic information into functional protein sequences. While the genome defines a static set of tRNA gene sequences, the functional tRNA pool depends on the expression and aminoacylation levels of different tRNA species, as well as many modifications that fine-tune tRNA activity.1,2 Dynamically regulated tRNA pools dictate the fidelity of the translation process and proteome integrity of a cell. Considering the central role of tRNA in translation regulation, investigating the cellular composition of tRNA pools provides a deeper understanding of the translational state of the cell. High-resolution sequencing-based methods for tRNA sequencing have provided transformative insights into the physiological impact of tRNA expression, modification, and aminoacylation levels on cellular stress and/or disease conditions.3–6 More recently, large-scale genome engineering efforts have also employed tRNA sequencing to recapitulate the translational state of engineered cells.

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Poster
Improving High-Throughput Automated NGS Library Generation in a Core Sequencing Lab

To answer genomic research needs for a faster, streamlined DNA prep workflow that delivers high quality libraries from a variety of sample types, New England Biolabs developed the NEBNext UltraExpress™ DNA Library Prep Kits which have straightforward protocols for all DNA inputs, either pre-sheared or enzymatically sheared. The workflow incorporates master mixed reagents, reduced incubation times, and fewer cleanup steps in comparison to previous kits. Thanks to an improved chemistry and a wide tolerance of inputs, the kit allows for the generation of libraries in under 2 hours and has a uniform protocol that streamlines preparation of diverse DNA sample inputs. With an automated infrastructure, this new kit is able to provide some of the most accessible, high-throughput, quality NGS library prep of DNA samples available.

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Poster
Automating KAPA EvoPlus library preparation kits on firefly®

For the first time, we demonstrate that the Roche KAPA EvoPlus and KAPA EvoPlus PCR-free DNA library preparation workflows can be successfully automated on firefly® to create Illumina-compatible NGS libraries comparable to those prepared manually. The combination of these technologies enables significant time savings in the laboratory for whole genome sequencing applications, whilst also mitigating the risk of repetitive strain injuries (RSI) posed by such a substantial number of manual pipetting steps.

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Poster
Automated Library Preparation Using Watchmaker DNA Library Prep Kit with Fragmentation on firefly®

With the rapid growth of Next Generation Sequencing (NGS), having a robust automation-friendly library preparation solution is a minimum requirement for the handling of precious samples across many applications This poster highlights TGen-North’s process to evaluate several library prep solutions and automation systems with the goal of transitioning from their current manual library prep to an automated workflow, using SPT Labtech’s firefly along with the Watchmaker DNA Library Prep Kit with Fragmentation.

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Poster
Miniaturization of DNA library preparation for Illumina sequencing using automated positive displacement liquid handling

Advances in next generation sequencing have resulted in huge increases in throughput with associated decreases in costs. As a result, the process of library preparation has become even more of a financial and time constraint to high throughput sequencing core facilities. Automation, in the form of liquid handling robots, has been able to alleviate some of these bottlenecks. This work describes the application of SPT Labtech’s mosquito® HV positive displacement nanoliter liquid handler to successfully implement the preparation of DNA libraries for Illumina sequencing at one tenth the volume of the original manual protocol.

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Poster
Transforming PCR protocols in pandemic response with automated non-contact liquid dispensing

During the COVID-19 pandemic, robust polymerase chain reaction (PCR) workflows were required to detect the Novel Coronavirus 2019 (Severe Acute Respiratory Syndrome Coronavirus 2: SARS-CoV-2) virus in clinical samples. Accurate detection of the virus in clinical samples allowed for rapid infection containment and subsequently alleviated the pressure on healthcare services during this time. Automation of these workflows has the benefit of not only increasing throughput, but also minimizing variability and human error particularly when dealing with precious reagents and limited sample. This application note describes the process by which the UKHSA testing megalab, the Rosalind Franklin laboratory, transitioned from the manufacturer recommended extraction plate preparation workflow for an ePCR/MAGMAX MVPII, to an automated workflow using SPT Labtech’s non-contact reagent dispenser, dragonfly® discovery, resulting in an 80-fold increase in the sensitivity of the assay.

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Application note
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