NEBNext® Sample Preparation for Next Generation Sequencing
Preparation of high quality libraries at high yield is a critical first step in next generation sequencing workflows. NEBNext® reagents are a series of highly pure reagents that facilitate sample preparation of DNA and RNA and target enrichment of DNA for next generation sequencing. NEBNext kits are carefully designed to improve yields and library diversity using a broad range of input amounts from a wide variety of sample types. For example, each reagent in the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645) has been reformulated, resulting in a several-fold increase in library yield with as little as 500 pg of input DNA.
The most recent technology addition to the NEBNext product line is NEBNext Direct® for target enrichment. This unique hybridization-based technology enables high sensitivity variant calling with the speed and precision previously only feasible using multiplex PCR-based approaches. The NEBNext Direct Cancer HotSpot Panel provides highly specific hybridization-based capture of 190 common cancer targets from 50 genes.
Use our tool, NEBNext Selector, to determine the best NEBNext products for you.
NEBNext® Sample Preparation for Next Generation Sequencing includes these areas of focus:
- Illumina® Library Preparation
- RNA for Illumina®
- DNA for Illumina®
- NEBNext® Ultra™ II DNA Library Prep
- NEBNext® Ultra™ II RNA Library Prep
- Ion Torrent™ DNA Library Preparation
- DNA for Ion Torrent™
- 454™ Library Preparation
- DNA for 454™
- RNA for 454™
- DNA Library Preparation
- RNA Library Preparation
- DNA Fragmentation
- NEBNext® dsDNA Fragmentase®
- FFPE DNA Repair
- NEBNext FFPE DNA Repair Mix
- NEBNext® Automation
FAQs for NEBNext® Sample Preparation for Next Generation Sequencing
Protocols for NEBNext® Sample Preparation for Next Generation Sequencing
- Appendix A for use with NEBNext Ultra II RNA First Strand Synthesis Module (E7771)
- Appendix A for use with Ultra™ II RNA and Ultra II Directional RNA (E7760, E7765, E7770, E7775)
- Guidelines for Running Samples on the Illumina MiSeq (E7000)
- Guidelines for Running Samples on the Illumina MiSeq (E6627)
- Guidelines for Setting up PCR Reactions (E6627)
- Guidelines for Setting Up PCR Reactions (NEB #E7000X only)
- Index Pooling Guidelines - 96 Single Index Kit (E6609)
- Index Pooling Guidelines (E6627)
- Index Pooling Guidelines (NEB #E7000)
- Protocol for a PCR reaction using NEBNext® Ultra™ II Q5® Master Mix (M0544)
- Protocol for Directional RNA-seq Workflows and NEBNext® Ultra™ II RNA First Strand Synthesis Module (E7771)
- Protocol for NEBNext Direct BRCA1/BRCA2 Panel (E6627)
- Protocol for NEBNext Direct® Cancer HotSpot Panel (NEB #E7000)
- Protocol for Non-directional RNA-seq Workflows and NEBNext® Ultra™ II RNA First Strand Synthesis Module (E7771)
- Protocol for use with FFPE RNA, NEBNext® rRNA Depletion Kit (Human/Mouse/Rat) (E6310) and NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (E7760, E7765)
- Protocol for use with FFPE RNA, NEBNext® rRNA Depletion Kit (Human/Mouse/Rat) (E6310) and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (E7770, E7775)
- Protocol for use with NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (E7645, E7103)
- Protocol for use with NEBNext® Ultra™ II End Repair/dA-Tailing Module (E7546)
- Protocol for use with NEBNext® Ultra™ II Ligation Module (E7595)
- Protocol for use with NEBNext® Poly(A) mRNA Magnetic Isolation Module (E7490) and NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (E7760, E7765)
- Protocol for use with NEBNext® Poly(A) mRNA Magnetic Isolation Module (E7490) and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (E7770, E7775)
- Protocol for use with NEBNext® rRNA Depletion Kit (Human/Mouse/Rat) (E6310) and NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (E7760, E7765)
- Protocol for use with NEBNext® rRNA Depletion Kit (Human/Mouse/Rat) (E6310) and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (E7770, E7775)
- Protocol for use with Purified mRNA or rRNA Depleted RNA and NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (E7760, E7765)
- Protocol for use with Purified mRNA or rRNA Depleted RNA and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (E7770, E7775)
- Protocol for use with rRNA Depleted FFPE RNA and NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (E7760, E7765)
- Protocol for use with rRNA Depleted FFPE RNA and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (E7770, E7775)
- Setting up the PCR Reactions < 96 Samples and 96 Samples - 96 Single Index Kit (E6609)
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NEBNext® for Illumina® Brochure
The NEBNext Brochure for Illumina provides an overview of library preparation for DNA, ChIP-Seq, RNA and Small RNA for the Illumina platform, as well as tech tips and select peer-reviewed citations.
- Addressing Challenges in Microbiome DNA Analysis
- The Quantitation Question: How does accurate library quantitation influence sequencing?
Other Tools & Resources
Brochures
Feature Articles
- Luck, A.N., Anderson, K.G., McClung, C.M., VerBerkmoes, N.C., Foster, J.M., Michalski, M.L. and Slatko, B.E. 2015. Tissue-specific transcriptomics and proteomics of a filarial nematode and its Wolbachia endosymbiont BMC Genomics. , PubMedID: 26559510, DOI:
Publications related to NEBNext® Sample Preparation for Next Generation Sequencing
Legal Information
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.
Videos
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NEBNext Direct® Target Enrichment
Learn about the innovative NEBNext Direct technology for target enrichment for next generation sequencing.
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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.
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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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Which NEBNext Product Format Should I Choose?
Not sure how sets and kits differ from modules? Ping explains the difference, and helps you to decide which one is right for you.
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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.
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Which DNA Library Prep Kit for Ion Torrent Should I Choose?
Having trouble deciding which NEBNext Fast DNA Library Prep Set for Ion Torrent™ to choose? Let Christine explain which set is right for your DNA.
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Tips for Bead-based Clean ups and Size Selection
Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.
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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.
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Optimization of NGS Library Preparation: Low Inputs and Fast, Streamlined Workflows
Learn how NEBNext® Ultra™ is enabling library prep for multiple applications, with lower input amounts and fast, streamlined workflows in this GenomeWeb webinar.
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A Breakthrough Method of Small RNA Sample Preparation
The common problem of adaptor dimer formation during small RNA library construction can be avoided by using NEBNext® protocols. Learn about this technique, and how it improves both performance and sensitivity in library construction.
