Library Preparation for Illumina

Illumina® sequencing instruments can be found in laboratories of all sizes, from lab-scale to production-scale, and NEBNext kits can be used to construct the high-quality libraries required for them all, for a wide array of sample types and applications.

NGS Library Preperation

The NEBNext® suite of products supports Illumina sequencing with sample preparation tools that streamline workflows, minimize inputs, and improve library yields and quality. NEBNext library preparation kits are available for both DNA library prep and RNA library prep; kits and modules are available for genomic DNA, FFPE DNA, methylated DNA, microbiome DNA, total RNA, single cell/low input RNA, and small RNA samples. These are complemented by a wide range of index primers and index adaptors for multiplexing. In addition to the extensive QCs performed on individual kit components, all NEBNext kits for Illumina library prep are functionally validated by preparation of a library, followed by Illumina sequencing. NEBNext library preparation reagents are available in multiple kit and workflow formats, for maximum convenience and flexibility. Designed for use in concert with NEBNext Multiplex Oligos for simple yet powerful NGS sample multiplexing.

Introducing: NEBNext UltraExpress™ library prep
Streamlined for speed.

The NEBNext UltraExpress library prep kits are the latest generation of library prep reagents for DNA or RNA, designed to be faster, simpler, and even more automation friendly. The new kits generate high-quality libraries from a variety of sample types with a single protocol for all inputs within the specified input range. The workflows incorporate master mixed reagents, reduced incubation times, and fewer cleanup steps, keeping hands-on time at a minimum.


NEBNext UltraExpress DNA Library Prep Workflow

NEBNext UltraExpress FS DNA Library Prep Kit Workflow

E3340 Workflow

NEBNext UltraExpress RNA Library Prep Kit Workflow


Savings* with NEBNext UltraExpress


Savings Infographic

* As compared to NEBNext Ultra II


NEBNext Ultra™ II is at the heart of our DNA and RNA library prep workflows.

The NEBNext Ultra II reagents and workflow are at the heart of NEB’s portfolio for next generation sequencing library preparation. NEBNext Ultra II kits and modules for Illumina® are an optimized combination of reagents and simplified workflows, enabling you to create DNA or RNA libraries of highest quality and yield, even when starting from extremely low or low-quality input amounts.

The Ultra II workflow is at the heart of: 

NEBNext Ultra II DNA Library Prep Workflow

Don’t just take our word for it; review the nearly 25,000 publications featuring NEBNext reagents, instead.




Choose Type:

Library Preparation for Illumina includes these subcategories:
DNA Library Prep for Illumina
RNA Library Prep for Illumina
FAQs for Library Preparation for Illumina
Application Notes for Library Preparation for Illumina
NEBNext UltraExpress™ DNA Library Prep Kit Workflow
Worklow image
NEBNext UltraExpress™ RNA Library Prep Kit Workflow
Workflow image
Single Cell/Low Input RNA Library Preparation Workflow for Illumina
Ultra II FS DNA Library Preparation Workflow
Ultra II Directional RNA Library Preparation Workflow for Illumina
Ultra II Non-directional RNA Library Preparation Workflow for Illumina
Ultra II DNA Library Preparation for Illumina
Small RNA Library Preparation Workflow for Illumina
Legal Information

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.


Give NEBNext a try with your own samples.

We offer samples of many of our NEBNext kits, modules and oligos.



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