The NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent® includes a master mix of reagents for enzymatic DNA fragmentation combined with end repair. This enables efficient and tunable DNA shearing and blunting with minimized sample loss before performing adaptor ligation and high-fidelity library amplification. The kit is compatible with input amounts from 10 ng - 1µg, with a workflow time of under 2 hours and ~12 min. hands-on time.
Adaptors and primers for singleplex libraries are supplied in the kits. For multiplexed libraries, the Ion Xpress™ Barcode Adapters from Thermo Fisher Scientific can be used.
If mechanical shearing of DNA is preferred, a version of the kit without enzymatic fragmentation reagents is available.
DNA Library Preparation Workflow for Ion Torrent
Representative Results from Libraries Constructed Using NEBNext for Ion Torrent
0.5 µg of DNA from 3 different genomes with varying GC content were used to construct 200 bp and 400 bp libraries using the NEBNext Fast DNA Fragmentation and Library Prep Set for Ion Torrent, analyzed by the Agilent® Bioanalyzer®.
Representative Ion Torrent Run Report
A typical Ion Torrent run report for libraries made from E. coli (K12 MG1655 strain) genomic DNA using NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent. A. Alignment Summary. 72 Mb of data was generated, with an average genome coverage of 15.5 X, from approximately 0.5 Million 200 bp reads. B. Read length histogram from 200 bp run.
Figure 1: Relative size distribution of Fragmented End Repaired DNA as seen using the Bioanalyzer® 2100 (Agilent Technologies, Inc.).
1 µg of E. coli DNA was fragmented and end repaired for 20 minutes at 25°C, followed by 10 minutes at 70°C.
Figure 2: Final Library size distribution using AMPure XP Bead Size Selection.
Figure 3: Final Library Size distribution using E-Gel Size Selection.
Lot Control
The lots provided are managed separately and qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page
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