DNA Library Prep for Illumina

New England Biolabs now offers two suites of reagents for DNA library prep, each solving a different set of challenges:

The NEBNext® Ultra™ II solution for Illumina DNA library prep is a streamlined suite of reagents and workflows designed for the construction of high-quality libraries, using input amounts from pg to μg of DNA. Workflows are fast, user-friendly, and automatable. Available with or without an integrated enzymatic fragmentation step and optional PCR-free workflows, NEBNext offers flexible product formats maximal flexibility in your workflows. Designed for use in concert with NEBNext Multiplex Oligos for simple yet powerful NGS sample multiplexing. 

The NEBNext UltraExpress™ reagents and workflow for DNA library prep are even more streamlined, enabling library prep in under 2 hours. With speed comes a tighter input amount range (10 – 200 ng), but for samples that fall within that input range, NEBNext UltraExpress offers a single-protocol solution; that means one adaptor concentration and one number of PCR cycles for all samples! And just like NEBNext Ultra II, our NEBNext UltraExpress DNA library prep kits are compatible with our NEBNext Multiplex Oligos.


NEBNext Ultra II DNA & FS DNA Library Prep

NEBNext UltraExpress DNA Library Prep


Unprecedented performance, enabling lower inputs and fewer PCR cycles for the broadest range of input amounts and qualities; input-to-library time: 5 hours

Streamlined and straightforward DNA library prep with a simple, single-condition protocol for all inputs within the kit’s input range; input-to-library time: < 2 hours

DNA Library Prep Kits:

NEBNext Ultra II DNA Library Prep Kit
With beads: NEB #E7103
Without beads: NEB #7645

NEBNext UltraExpress DNA Library Prep Kit
Without Beads: NEB #E3325

Pre-sheared Input Amount

500 pg – 1 µg of sheared DNA

10 ng – 200 ng of sheared DNA

Modules available?


FS DNA Library Prep Kits:

NEBNext Ultra II FS DNA Library Prep Kit
With beads: NEB #E6177
Without beads: NEB #E7805


Intact Input Amount

100 pg – 500 ng of intact DNA


Modules available?

Ultra II FS DNA Module NEB #E7810)


Compatible with PCR-free workflows?

Yes. NEBNext Ultra II DNA PCR-free and FS DNA PCR-free library prep kits are available

Data coming soon

Compatible with FFPE DNA?

For FFPE DNA library prep, we strongly recommend the NEBNext UltraShear FFPE DNA Library Prep Kit. We do not recommend use of NEBNext FS DNA kits for FFPE DNA.

Data coming soon

Compatible with methylome analysis?

We strongly recommend NEBNext EM-seq™ (NEB #E7120) for methylome analysis, however NEBNext DNA kits can be used. We do not recommend use of NEBNext FS DNA kits with methylated DNA due to the potential for loss of methyl marks.

Compatible with oxford nanopore technologies?

No. We recommend the NEBNext Companion Module for Oxford Nanopore Technologies® Ligation Sequencing (NEB #E7180).

NEBNext DNA library prep reagents are also available at scale for your high-throughput and/or development needs. Contact the Customized Solutions Team for more details on custom product options, large volumes, or OEM materials.  

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FAQs for DNA Library Prep for Illumina
Related Categories
NEBNext UltraExpress™ DNA Library Prep workflow
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Ultra II FS DNA Library Preparation Workflow
ChIP DNA Library Preparation Workflow for Illumina
Ultra II DNA Library Preparation for Illumina
DNA Product Details for Illumina
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