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Illumina® Library Preparation

Library preparation for the Illumina sequencing platform requires fragmentation of DNA, or use of cDNA prepared from RNA, followed by repair of 3’ and 5’ ends to form blunt-ended, phosphorylated molecules, and the addition of a non-templated dA-tail before ligation to an adaptor. If necessary to achieve sufficient yields, the final step is PCR amplification of the library. Small RNA libraries are constructed using a different workflow, in which adaptors are ligated directly to the small RNA molecules, followed by reverse transcription and PCR amplification. For more detail, select a workflow tab below.

The NEBNext suite of products supports Illumina sequencing with sample preparation tools that streamline workflows, minimize inputs, and improve library yields and quality. NEBNext sample preparation kits are available for genomic DNA, ChIP DNA, FFPE DNA, Microbiome DNA, RNA and small RNA samples. In addition to the extensive QCs performed on individual kit components, all NEBNext kits for Illumina 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.

GenomeWeb article reviews NEBNext Small RNA Kits.

 

Illumina® is a registered trademark of Illumina, Inc.
NEBNext® is a registered trademark of New England Biolabs, Inc.

Illumina® Library Preparation includes these areas of focus:

RNA for Illumina®
DNA for Illumina®
NEBNext® Ultra™ II DNA Library Prep
NEBNext® Ultra™ II RNA Library Prep

FAQs for Illumina® Library Preparation

Protocols for Illumina® Library Preparation

Ultra II FS DNA Library Preparation Workflow

Ultra II Directional RNA Library Preparation Workflow for Illumina

Workflow"

Ultra II  Non-directional RNA Library Preparation Workflow for Illumina

Workflow"

Ultra II DNA Library Preparation Workflow for Illumina

Ultra DNA Library Preparation Workflow for Illumina

Small RNA Library Preparation Workflow for Illumina

Legal Information

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

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.

  1. NEBNextUltra2DirectionalRNA_720

    NEBNext Ultra II Directional RNA Workflow

    Learn more about the streamlined workflow for the NEBNext Ultra II Directional RNA Library Prep Kit.

  2. Size Selection and Cleanup with NEBNext Ultra

    Size Selection and Cleanup with NEBNext Ultra II and SPRI beads

    This video walks you through the size selection and cleanup steps using the NEBNext Ultra II DNA

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    Tips for Optimizing DNA Inputs in NGS Library Construction

    Optimizing DNA inputs for NGS library prep is an important step, if you want to ensure a high-quality library. Start by following these tips.

  4. NEBNextVideo_OptimizingRNAInput_thumb

    Tips for Optimizing RNA Inputs in NGS Library Construction

    Watch as Deyra explains how to optimize your RNA inputs for successful NGS library preparation.

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    NEBNext Ultra II DNA Library Prep Protocol

    This video walks you through DNA Library Preparation using the NEBNext Ultra II DNA Library Prep Kit. The video also includes tips for optimization as well as safe stopping points.

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    12 Quick Tips for NGS Library Preparation

    Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.

  7. NEBNextVideo_CoreLabFacilities_thumb

    NEBNext for Core Lab Facilities

    Are you running a core sequencing facility? Watch as Eileen shares a few reasons that NEBNext products are particularly well suited to sequencing core labs.

  8. Behind the paper: DNA damage is a pervasive cause of sequencing errors, directly confounding variant identification

    Behind the paper: DNA damage is a pervasive cause of sequencing errors, directly confounding variant identification

    NEB researchers published a paper in Science highlighting DNA damage as a prevalent source of errors in public cancer databases. Learn about how addressing this damage could improve the detection of low-frequency disease variants.

  9. NEBTV Webinar

    How Library Preparation Affects Sequencing Accuracy

    In this webinar, Laurence Etwiller and Jennifer Ong discuss their recent publications, and how library preparation affects sequencing accuracy and variant calling. View full Q & A summary.