High-quality, high-yield library preparation is a critical first step towards optimizing your next generation sequencing (NGS) workflows. The NEBNext® product line is a series of highly pure reagents for both DNA and RNA sample prep, as well as target enrichment of DNA for NGS. NEBNext kits are carefully designed and QC’d to improve yields and library diversity, using a broad range of input amounts from a wide variety of sample types. Recently, a novel enzyme-based DNA fragmentation system was added to enable streamlined DNA fragmentation, end repair, and dA tailing with a single enzyme mix; this kit, the NEBNext Ultra II FS DNA Library Prep Kit (NEB #E7805), enables reliably high-quality and high-yield library construction with as little as 100 pg of input DNA.
For those looking for a more targeted approach, NEBNext provides NEBNext Direct® for target enrichment. This unique hybridization-based technology enables high-sensitivity variant calling with a speed and precision previously only feasible using multiplex PCR-based approaches. For convenient customization, NEBNext Direct Custom Ready Panels offer ready-made, optimized baits for over 850 targets, enabling faster time to targeted results. For pre-selected disease-relevant panels, the NEBNext Direct Cancer HotSpot Panel (NEB #E7000) provides highly specific hybridization-based capture of 190 common cancer targets from 50 genes, while the NEBNext Direct BRCA1/BRCA2 Panel (NEB #E6627) generates 100% coverage of all protein coding regions in BRCA1 and BRCA2 genes.
- DNA Fragmentation
- DNA Library Preparation
- FFPE DNA Repair
- NEBNext FFPE DNA Repair Mix
- Illumina® Library Preparation
- NEBNext® Ultra™ II RNA Library Prep
- NEBNext® Ultra™ II DNA Library Prep
- RNA for Illumina®
- DNA for Illumina®
- Ion Torrent® DNA Library Preparation
- DNA for Ion Torrent®
- NEBNext® Automation
- RNA Library Preparation
- Protocol for use with NEBNext® Ultra™ II End Repair/dA-Tailing Module (E7546)
- Protocol for use with NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (E7645, E7103)
- Protocol for a PCR reaction using NEBNext® Ultra™ II Q5® Master Mix (M0544)
- Setting up the PCR Reactions < 96 Samples and 96 Samples - 96 Single Index Kit (E6609)
- Index Pooling Guidelines - 96 Single Index Kit (E6609)
- Protocol for NEBNext Direct® Cancer HotSpot Panel (NEB #E7000)
- Guidelines for Setting Up PCR Reactions (E7000X only)
- Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490)
- Protocol for use with FFPE RNA, NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
- Protocol for use with NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765)
- Protocol for use with Purified mRNA or rRNA Depleted RNA and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765)
- Protocol for use with rRNA Depleted FFPE RNA and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765)
- Protocol for use with NEBNext® UltraTM II RNA Library Prep Kit for Illumina® E7770 and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
- 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 FFPE RNA, NEBNext Ulta II RNA Library Prep Kit for Illumina E7770 and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
- Protocols for use with Purified mRNA or rRNA Depleted RNA and NEBNext Ultra II RNA Library Prep Kit for Illumina
- Protocol for use with rRNA Depleted FFPE RNA and NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB # E7770, #E7775)
- Guidelines for Running Samples on the Illumina MiSeq (E7000)
- Protocol for Directional RNA-seq Workflows and NEBNext Ultra II RNA First Strand Synthesis Module (E7771)
- Protocol for Non-directional RNA-seq Workflows and NEBNext Ultra II RNA First Strand Synthesis Module (E7771)
- Appendix A
- Appendix A for use with NEBNext Ultra II RNA First Strand Synthesis Module (E7771)
- Guidelines for Running Samples on the Illumina MiSeq (E6627)
- Guidelines for Setting up PCR Reactions (E6627X only)
- Protocol for NEBNext Direct BRCA1/BRCA2 Panel (E6627)
Addressing Challenges in Microbiome DNA Analysis
Microbiome sample analysis is commonly confounded by the presence of host-cell DNA in the sample. The NEBNext® Microbiome DNA Enrichment Kit enables enrichment of non-CpG-methylated DNA from samples contaminated with eukaryotic-cell DNA.
Improving Enzymatic DNA Fragmentation for Next Generation Sequencing Library Construction
Among the available options for DNA fragmentation, enzymatic fragmentation is demonstrably gentler on the sample, yielding less damage at any scale. The new NEBNext® Ultra™ II FS DNA Library Prep Kit for Illumina® includes a single step for enzymatic fragmentation, end repair, and dA tailing.
The Quantitation Question: How does accurate library quantitation influence sequencing?
The determination of the number of sequencing-ready molecules present after library preparation is an important step in the next generation sequencing (NGS) workflow and has a strong influence on the success of both a sequencing run and a sequencing-based experiment.
- A Robust, Streamlined, Enzyme-based DNA Library Preparation Method Amenable to a Wide Range of DNA Inputs (2019)
- A Single-tube, Low Input Protocol for Long Read Sequencing (2019)
- AGBT 2018 Wellcome Trust Sanger Institute poster: Thurston. S., et al. (2018). A Large Genome Centre Core Pipeline Refresh.
- Application of a Novel, Targeted Sequencing-Based Genotyping Approach for Cost Effective Marker Assessment in O. sativa (2019)
- Benefits of Illumina® Library Quantitation with NEBNext® Library Quant Kit (2015)
- EM-seq™ Enables Accurate and Precise Methylome Analysis of Challenging DNA Samples (2019)
- Enzymatic Methyl-seq: Next Generation Methylomes (2019)
- Examining Sources of Error in PCR by Single-Molecule Sequencing (2017)
- High Quality Automation Libraries Prepared from a Broad Range of DNA Input Amounts (2017)
- High-Throughput Screening of 2300 Genetic Markers in S.lycopersicum using the NEBNext Direct Multiplexed Genotyping Approach (2019)
- Improving Transcriptome Profiling for Single Cell and Low Input RNA (2019)
- NEBNext Direct® Custom Ready Panels Overcome Challenges Associated with Targeted Re-sequencing (2019)
- NEBNext® Ultra™II FS DNA: A Robust Enzyme-based DNA Library Preparation Method Compatible with Plant and Animal Samples (2019)
- Uncovering the Cannabis sativa Methylome Through Enzymatic Methyl-seq (2019)
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 [email protected].
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.
Learn more about the streamlined workflow for the NEBNext Ultra II Directional RNA Library Prep Kit.
This video walks you through the size selection and cleanup steps using the NEBNext Ultra II DNA
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 for DNA quantification and assessing purity.
Watch as Deyra explains how to optimize your RNA inputs for successful NGS library preparation.
View our tutorial for help visualizing NEBNext Direct’s unique and enabling workflow.
Learn about the innovative NEBNext Direct technology for target enrichment for next generation sequencing.
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.
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.
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.
These 12 quick tips for NGS library preparation are a great refresher if you're a seasoned pro, or a great introduction if you're a beginner.