Work with University of California, San Diego, has resulted in a new workflow for single cell sequencing that reduces the cost of sample preparation by 90%
A collaboration between UK company TTP Labtech, a global leader in the design and development of automated instrumentation and consumables for life science applications, and the University of California San Diego (UC San Diego) has resulted in a new workflow for single cell sequencing that reduces the cost of sample preparation by 90%.
The workflow, which has been published in the Journal of Laboratory Automation, was developed through the combination of UC San Diego’s expertise in genomics and the use of TTP Labtech’s mosquito liquid handlers, which can pipette nanolitre to microlitre volumes with high accuracy and reproducibility, to miniaturise and automate the process.
Although single-cell genomics is advancing rapidly, researchers face challenges linked to reproducibility, sensitivity, scalability and cost, particularly when trying to miniaturise high-throughput applications. To maximise accuracy and precision, library prep protocols recommend volumes that are within the range of manual pipettes or large-volume liquid handlers. However, only a small proportion of each of the libraries prepared will be required for sequencing.
The focus of UC San Diego’s partnership with TTP Labtech was to miniaturise single-cell RNA library prep reaction set-up, to minimise sample and reagent requirements, thereby reducing library prep costs while maintaining throughput, accuracy and reproducibility.
The miniaturisation of library prep volumes was achieved using TTP Labtech’s mosquito HTS and HV liquid handlers, which accurately dispense volumes of between 25 nL and 1.2 µL and 0.5 to 5 µL, espectively. Both instruments use true positive-displacement pipetting technology, which maintains volumetric accuracy even for highly viscous liquids, enabling high concentration gDNA to be handled, to save on reagent cost and sample input. To assess the quality of the libraries, UC San Diego applied the workflow to analyse pancreatic differentiation of human embryonic stem cells. The resulting single-cell RNA-sequencing data were analysed to determine the reproducibility of this system and its ability to distinguish not only between cells at different stages of differentiation but also between individual cells within each stage.
'Single cell sequencing requires large numbers of samples to be handled to provide valid data based on population studies,' said Joby Jenkins, Product Manager, TTP Labtech. 'Reproducibility and sensitivity, as well as scalability and cost, can be limiting factors when large numbers of cells are analysed, and we are very pleased that our partnership with UCSD has achieved our goal of improving sample prep for single cell genomics research.'
Dr Louise Laurent, Associate Professor, UC San Diego, added: 'The workflow we have developed, in partnership with TTP Labtech, has the potential to provide researchers with significant cost savings while maintaining the quality of single cell sequencing. The instrument cost and their ease-of-use make the mosquito liquid handlers viable options for labs that would like to scale-down and automate next-generation sequencing sample preparation.'