RESUMEN
Next generation sequencing is in the process of evolving from a technology used for research purposes to one which is applied in clinical diagnostics. Recently introduced high throughput and benchtop instruments offer fully automated sequencing runs at a lower cost per base and faster assay times. 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. Library preparation protocols usually consist of a multistep process and require costly reagents and substantial hands-on-time. Considerable emphasis will need to be placed on standardisation to ensure robustness and reproducibility. This review presents an overview of the current state of automation of library preparation for next generation sequencing. Major challenges associated with library preparation are outlined and different automation strategies are classified according to their functional principle. Pipetting workstations allow high-throughput processing yet offer limited flexibility, whereas microfluidic solutions offer great potential due to miniaturisation and decreased investment costs. 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. Finally, two applications, the characterisation of bacterial pathogens and the sequencing within human immunogenetics, are outlined and benefits of automation are discussed.
Asunto(s)
Secuenciación de Nucleótidos de Alto Rendimiento , ARN , Automatización , Biblioteca de Genes , Humanos , Reproducibilidad de los ResultadosRESUMEN
Due to the fact that the use of iodated table salt is not compulsory in Germany and entirely at the discretion of the individual consumer, there is hardly any effective prevention of endemic iodine deficiency struma in this country. The good results achieved in the former GDR with iodine prophylaxis dropped to the FRG level after the unification. Using iodated salt when preparing individual food dishes is not enough to counteract iodine deficiency. We proved this in our study which is communicated below. An iodine deficiency of the first degree remains. Effective compensation of iodine deficiency can be achieved only if iodine is added on a large scale in all branches of food preparation (including the serving and catering of food), of the food industry (on both a major and a small scale) and in meat production, i.e. in producing food of animal origin.