Next-generation sequencing library preparation: simultaneous fragmentation and tagging using in vitro transposition

DNA is fragmented and the transferred strand of the transposon end oligonucleotide is covalently attached to the 5′ end of the target fragment (Fig. 1a). The size distribution of the fragments can be controlled At present, next-generation sequencing platforms use slightly different technologies for sequencing, such as pyrosequencing, sequencing by synthesis or sequencing by ligation. However, most platforms adhere to a common library preparation procedure, with minor modifications, before a ‘run’ on the instrument. This procedure includes fragmenting the DNA (sonication, nebulization or shearing), followed by DNA repair and end polishing (blunt end or A overhang) and, finally, platform-specific adaptor ligation. This process typically results in considerable sample loss with limited throughput. To streamline the workflow, increase throughput and reduce sample loss, Epicentre has developed NexteraTM technology, a transposon-based method for preparing fragmented and tagged DNA libraries in as little as 4 hours. This flexible, scalable and efficient technique can be used to generate libraries for multiple sequencing platforms.