Advances in next-generation sequencing have led to new library preparation methods compatible with multiple sequencing platforms. Current methods (both mechanical and enzymatic) face limitations: multi-step protocols, sample loss, lack of automation and labor costs. With the continued decline of sequencing costs and increase in sample throughput, there is greater demand for more costeffective, streamlined library preparation methods. Here we describe Epicentre's patented Nextera ™ technology, which addresses current issues in sample preparation and provides a simplified procedure amenable to high-throughput workflows.
Molecular weight distribution of Nextera-generated librariesThe molecular weight distributions of libraries prepared using Nextera technology can be controlled for the read-length requirements of different sequencers. To show that molecular weight distributions are consistent and reproducible across different sample types, we prepared genomic DNA libraries from phage λ, Escherichia coli and human DNA using both Roche 454-Compatible and Illumina-Compatible Nextera Enzyme Mixes according to the standard protocol (Fig. 1). The reaction conditions were optimized to yield fragment sizes appropriate for the respective sequencers. Each final fragment size includes approximately 100 base pairs (bp) of adaptor sequence; therefore, the actual genomic DNA sequenced is smaller than the apparent molecular weight of the library. It should also be noted that not all sample types will result in similar molecular weight distributions, as many factors (for example, complexity of the sample type and quantification method) can affect the molecular weight distribution.
Deep sequencing of Nextera Roche 454-compatible librariesLibraries were prepared from multiple sample types and then sequenced to validate Nextera technology for Roche 454. E. coli, plasmid, cosmid and soy genomic DNA was fragmented and 5′ end-tagged with Nextera Enzyme Mix (Roche-Compatible). Roche
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.