Library preparation is a critical part of the next generation sequencing workflow; successful sequencing requires the generation of high quality libraries of sufficient yield and quality.As sequencing technologies improve and capacities expand, boundaries are also being pushed on sample preparation. High performance is required from ever-decreasing input quantities and from samples of lower quality. At the same time, the need is increasing for faster, automatable protocols that perform reliably and do not compromise the quality of the libraries produced.
To meet these growing challenges, the NEBNext suite of products continues to evolve to support next generation sequencing with sample preparation tools that streamline workflows, minimize inputs, and improve library yields and quality.
NEBNext reagents are available for sample preparation for Illumina®, IonTorrent®, and Oxford Nanopore Technologies® (ONT) sequencing platforms. Products are in user-friendly formats including kits and modules, with bulk or customized formats also available. Adaptors and primers are available separately, for maximized flexibility. Use of NEBNext products has been cited in over 11,000 publications.
- DNA Library Prep – NEBNext DNA library prep kits utilize fast, streamlined workflows and novel reagents that have been designed for performance. Sample prep kits are available for a broad range of input amounts, from pg to μg of DNA.
- FFPE DNA – NEBNext offers two options to addresses the challenges of FFPE DNA: repair of DNA damage caused by FFPE treatment and high-efficiency library prep.
- Multiplex Oligos (Adaptors & Primers) – NEBNext Multiplex Oligos are a selection of index primers and index adaptors in a variety of configurations for maximal flexibility in the library prep workflow.
- ChIP-Seq Library Prep – The low input amounts typical of ChIP-Seq library prep are compatible with NEBNext high-efficiency, streamlined workflows.
- RNA Library Prep – NEBNext products are available for directional and non-directional RNA library prep with novel reagents and streamlined workflows, designed for high quality library generation even from low nanogram quantities of RNA. High-performance rRNA depletion and mRNA enrichment products are also available.
- Target Enrichment – The novel NEBNext Direct® technology uses a unique approach to hybridization-based target enrichment, and generates high quality libraries in a single day. NEBNext library prep kits are compatible with standard target enrichment protocols.
- Microbiome DNA Enrichment – NEBNext products enable separation of microbiome DNA from contaminating host DNA, as well as DNA library preparation from low input amounts.
- Library Quantitation – The NEBNext qPCR-based Library Quant Kit enables accurate and reproducible quantitation of Illumina libraries, with a user-friendly workflow.
One or more of these products are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc.
Illumina® is a registered trademark of Illumina, Inc. Ion Torrent™ is a trademark of Life Technologies, Inc.
- Protocol for NEBNext Ultra II DNA Library Prep with Sample Purification Beads (E7103)
- 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)
- 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)
- Protocol for a PCR reaction using NEBNext® Ultra™ II Q5® Master Mix (M0544)
- Protocol for use with NEBNext® Ultra™ II End Repair/dA-Tailing Module (E7546)
- Setting up the PCR Reactions < 96 Samples and 96 Samples - 96 Single Index Kit (E6609)
- Index Pooling Guidelines - 96 Single Index Kit (E6609)
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.
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.
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.
Enzymatic Methyl-seq: The Next Generation of Methylome Analysis
- NEBNext for Ion Torrent™ Brochure
- NEBNext® for DNA Sample Prep
- The NEBNext Sample Preparation Workflow Poster
- NEBNext Custom RNA Depletion Design Tool
- Benefits of Illumina® Library Quantitation with NEBNext® Library Quant Kit (2015)
- High Quality Automation Libraries Prepared from a Broad Range of DNA Input Amounts (2017)
- AGBT 2018 Wellcome Trust Sanger Institute poster: Thurston. S., et al. (2018). A Large Genome Centre Core Pipeline Refresh.
- Examining Sources of Error in PCR by Single-Molecule Sequencing (2017)
- 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)
- Application of a Novel, Targeted Sequencing-Based Genotyping Approach for Cost Effective Marker Assessment in O. sativa (2019)
- EM-seq™ Enables Accurate and Precise Methylome Analysis of Challenging DNA Samples (2019)
- Enzymatic Methyl-seq: Next Generation Methylomes (2019)
- 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)
- A highly multiplexed target enrichment approach for sample identification and tracking using the NEBNext Direct Genotyping Solution
- EM-seq™ enables accurate and robust methylation detection of cell free DNA and FFPE DNA sample types
- Enzymatic Methyl-seq: Next Generation Methylomes
- Increasing Power to Detect Low-Frequency Variants Using Dual-Unique Molecular Indices
- Increasing Sensitivity of Transcriptome Profiling in Prokaryotic and Eukaryotic Samples by Depleting Abundant RNAs
- NEBNext® Ultra™ II FS DNA: A Robust Enzyme-based DNA Library Preparation Method Compatible with Plant Samples
- An improved multiplex targeted amplicon sequencing of SARS-CoV-2 using Oxford Nanopore Technology
- Enzymatic Methyl-seq enables accurate and robust methylation detection
- Improving multiplex targeted amplicon sequencing of SARS-CoV-2 on Illumina platforms
- Improving transcriptome analysis by incorporating Unique Molecular Identifiers in RNA-sequencing libraries
- Incorporation of Unique Molecular Identifiers (UMIs) into Unique Dual Indexing (UDI) of samples improves the accuracy of quantitative Next Generation Sequencing
- Increasing the sensitivity of transcriptome profiling in eukaryote and blood samples by depleting abundant RNAs
- Primer-monitor.neb.com – streaming detection and notification of new SARS-CoV-2 variants
- Selective removal of abundant RNAs enhances the sensitivity of transcript detection across different prokaryotic and archaeabacterial species
- Induro™, a Novel Reverse Transcriptase for Nanopore Direct RNA Sequencing with Significantly Improved RNA 5′ Coverage
- Excellent performance – NEBNext continues to raise the bar on construction of high quality libraries from ever-decreasing input quantities, with a focus on high yields, high library diversity and minimized bias.
- Quality you can rely on – In addition to the extensive QCs performed on individual kit components, all NEBNext kits are functionally validated by preparation of a library, followed by sequencing. Use of NEBNext products has been cited in over 1,000 publications.
- Ease of use – NEBNext kits employ streamlined workflows, with minimized hands-on times and simple to follow protocols.
- Convenient product formats –
- NEBNext Kits include reagents for the entire library preparation workflow.
- Modules provide reagents for individual steps in a workflow and can be combined to cover an entire library prep workflow, or can be used in conjunction with other reagents to enable a customized workflow for your specific needs.
- Adaptors and primers are supplied separately from the NEBNext kits (as NEBNext Oligos modules), allowing for increased flexibility.
- SPRIselect® beads are provided as an optional component of our latest NEBNext Ultra™ II DNA Library Prep Kit for Illumina.
- Wide range of sample types and applications – NEBNext products are available for DNA, RNA, Small RNA, ChIP DNA, FFPE DNA and Microbiome DNA, for steps throughout the sample preparation workflow, from upstream enrichment to final library quantitation.
- Value pricing
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 email@example.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.
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.
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.
Get a quick overview of NGS library preparation from an NEB scientist.