New England Biolabs (NEB), Ipswich, Mass, has launched the NEBNext Ultra II FS DNA library prep kit for next-generation sequencing (NGS). The kit employs a novel enzymatic fragmentation system that combines fragmentation, end repair, and dA-tailing in a single tube, and utilizes a single protocol regardless of input amount or guanine-cytosine (GC) content.
“As applications for NGS continue to expand and sequencing instrument capacity continues to grow, laboratories must resolve the challenges of quickly preparing high-quality libraries from ever-decreasing input amounts in a scalable and user-friendly manner,” says Fiona Stewart, NEBNext portfolio manager at NEB.
“With the end user in mind, we’ve developed a new fragmentation technology that doesn’t require the conventional cleanup steps that can result in sample loss,” says Stewart. “The method is also extremely robust—the same protocol is used for all input DNA amounts and GC contents, and with input DNA in water, TE, or Tris. Fragmentation is further integrated into the library prep workflow by combining fragmentation, end repair, and dA-tailing reagents into a single mix, enabling subsequent adaptor ligation without a cleanup step.
“The combination of these high-reaction efficiencies and reduction in sample losses increases library yields and enables use of a wide range of input amounts, from 100 pg to 0.5 µg of eukaryotic DNA,” says Stewart. “Critically, the quality of the fragmentation system libraries is high, with excellent sequencing metrics and GC coverage uniformity, including with polymerase-chain reaction (PCR)-free libraries.”
With the kit’s streamlined workflow, library preparation can be completed within 2.5 hours, with less than 15 minutes of hands-on time. High-efficiency Ultra II ligation reagents are provided for adaptor ligation, and while the kit is compatible with PCR-free workflows with inputs as low as 50 ng of eukaryotic DNA, the NEBNext Q5 Ultra II PCR Master Mix is provided for amplification of libraries when necessary. The kit is also available with the option of SPRISelect beads for size-selection and clean-up steps.
For more information, visit New England Biolabs.