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 sequence-based target genotyping applications in plant, animal and human, the NEBNext Genotyping Solution enables pre-capture pooling of up to 96 samples for cost-effective and scalable genotyping of up to 5,000 genomic markers.
- 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® ARTIC products for SARS-CoV-2 sequencing
- NEBNext® Automation
- RNA Library Preparation
- Protocol for use with NEBNext® Ultra™ II End Repair/dA-Tailing Module (E7546)
- 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 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)
- Improved methods for site-directed mutagenesis using Gibson Assembly® Master Mix
- Improving NGS library performance with lower input amounts using the NEBNext Ultra II RNA Library Prep Kit for Illumina (non-directional) – Advances in non-strand-specific RNA library construction in a ribosomal RNA depletion workflow
- Increased transcription detection with the NEBNext Single Cell/Low Input RNA Library Prep Kit
- Selective Depletion of Abundant RNAs to Enable Transcriptome Analysis of Low-input and Highly-degraded RNA from FFPE Breast Cancer Samples
- AppNote NEBNext Library Quant Kit
- Improving NGS library performance with lower input amounts using the NEBNext Ultra II RNA Library Prep Kit for Illumina (non-directional) – Advances in non-strand-specific RNA library construction in a poly(A) mRNA enrichment workflow
- Obtain superior NGS library performance with lower input amounts using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina
- Monarch HMW DNA Extraction Kit improves sample preparation for Oxford Nanopore Technologies sequencing of malaria parasites
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.
- The NEBNext Sample Preparation Workflow Poster
- 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 Customizable Approach for Selective Removal of Abundant RNAs Enhances the Sensitivity of Transcript Detection Across Species
- 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
- Comparison of different sequencing platforms using 480 unique dual indexed libraries for 4 different applications
- 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
- Vladimir Potapov, Jennifer L. Ong. 2017. Examining Sources of Error in PCR by Single-Molecule Sequencing. PLOS One. , PubMedID: 28683110, DOI:
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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.
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Learn more about the streamlined workflow for the NEBNext Ultra II Directional RNA Library Prep Kit.
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View our tutorial for help visualizing NEBNext Direct’s unique and enabling workflow.
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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.
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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.