For the emerging field of single cell transcriptomics for example, microfluidics enable singularisation of tens of thousands of cells in nanolitre droplets and barcoding of the RNA to assign each nucleic acid sequence to its cell of origin.
Pipetting workstations allow high-throughput processing yet offer limited flexibility, whereas microfluidic solutions offer great potential due to miniaturisation and decreased investment costs. Major challenges associated with library preparation are outlined and different automation strategies are classified according to their functional principle. This review presents an overview of the current state of automation of library preparation for next generation sequencing. Considerable emphasis will need to be placed on standardisation to ensure robustness and reproducibility. Library preparation protocols usually consist of a multistep process and require costly reagents and substantial hands-on-time.
In turn, the complex and cumbersome library preparation, starting with isolated nucleic acids and resulting in amplified and barcoded DNA with sequencing adapters, has been identified as a significant bottleneck. Recently introduced high throughput and benchtop instruments offer fully automated sequencing runs at a lower cost per base and faster assay times. Next generation sequencing is in the process of evolving from a technology used for research purposes to one which is applied in clinical diagnostics.
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