Application Notes

Use this page to access application and technical notes available from NEB. View the full list below, or filter by application for a more targeted selection.

Filter by Application




73 Results


Optimizing rapid isothermal workflow for SARS-CoV-2 with WarmStart® LAMP with UDG

Last Updated November 2021

This application note describes the use of WarmStart Multi-purpose LAMP/RT LAMP 2X Master Mix (with UDG) for rapid and sensitive detection of SARS-CoV-2 RNA.

Read App Note


Obtain superior NGS library performance with lower input amounts using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina®

Last Updated October 2021

Application: NGS Sample Prep and Target Enrichment

This technical note describes RNA library preparation using a rRNA depletion workflow and the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina.

Read App Note


Automated library preparation using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® on the epMotion® 5075t

Last Updated October 2021

Application: NGS Sample Prep and Target Enrichment

This application note describes a robust automated workflow for the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina on the epMotion 5075t.

Read App Note


NEBExpress® Cell-free E. coli Protein Synthesis System

Last Updated October 2021

Application: Protein Expression and Purification

This technical note describes NEB’s new, versatile, lysate-based, cell-free protein synthesis system for use with circular or linear DNA, or mRNA templates.

Read App Note


Improved methods for site-directed mutagenesis using NEBuilder® HiFi DNA Assembly Master Mix

Last Updated September 2021

Application: Cloning and Synthetic Biology

This application note descibes the use of the NEBuilder® HiFi DNA Assembly Master Mix for the generation of single or multiple site-directed mutagens.

Read App Note


Protocol for using Recombinant Cas9 Nuclease to Assess Locus Modification in Genome Editing Experiments

Last Updated September 2021

Application: Cloning and Synthetic Biology

This application note describes sensitive in vitro digestion of PCR amplicons using Cas9 Nuclease for detecting indels in genome editing experiments.

Read App Note


A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers

Last Updated August 2021

Application: Cloning and Synthetic Biology, RNA Analysis

This technical note provides detailed explanations and guidance on how to interpret and use MVS measurements, with a focus on samples purified using silica spin columns.

Read App Note


Optimized Integration of New England Biolabs® Loop-mediated Isothermal Amplification (LAMP) Reagents with Axxin ISO Instruments

Last Updated August 2021

This app note details the optimized settings needed to analyze single and multiplex LAMP reactions on Axxin instruments.

Read App Note


Analysis of 5-methylcytosine and 5-hydroxymethylcytosine Levels in Human Brain and Liver DNA Samples using the EpiMark® 5-hmC and 5-mC Analysis Kit

Last Updated August 2021

Application: Epigenetics and Cellular Analysis

This application note demonstrates the use of the EpiMark 5-hmC and 5-mC Analysis Kit on different tissue samples.

Read App Note


Protein Expression with T7 Express Strains

Last Updated July 2021

Application: Glycobiology and Protein Analysis, Protein Expression and Purification

This application note provides protocols for T7 protein expression with NEB's T7 Express Competent E. coli.

Read App Note


Analysis of a Fusion Protein using the Protein Deglycosylation Mix II and Mass Spectrometry

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This application note describes the use of Protein Deglycosylation Mix II with PNGase F to show the presence of N- or O-glycans in the protein of interest.

Read App Note


Enzymatic removal of N- and O-glycans using PNGase F or the Protein Deglycosylation Mix

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This application note describes the enzymatic removal of glycans from a protein using PNGase F, or the Protein Deglycosylation Mix.

Read App Note


High-yield, Scalable Library Preparation with the NEBNext® Ultra™ II FS DNA Library Prep Kit

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This technical note describes DNA library preparation with enzymatic DNA fragmentation using the NEBNext Ultra II FS DNA Library Prep Kit.

Read App Note


Labeling of Escherichia coli Expressed SNAP-tag® Fusion Proteins

Last Updated July 2021

Application: Epigenetics and Cellular Analysis, DNA Amplification PCR and qPCR, Glycobiology and Protein Analysis

This application note provides protocols for labeling of E. coli expressed SNAP-tag fusion proteins.

Read App Note


Detailed Characterization of Antibody Glycan Structure using the N-Glycan Sequencing Kit

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This application note describes the NEB N-glycan Sequencing Kit for the characterization of glycans on Fc domains of therapeutic antibodies and fusion proteins.

Read App Note


Characterization of Glycans from Erbitux®, Rituxan® and Enbrel® using PNGase F (Glycerol-free), Recombinant

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This application note describes the characterization of glycans from Erbitux, Rituxan and Enbrel using PNGase F (Glycerol-free), Recombinant.

Read App Note


Unbiased and fast IgG deglycosylation for accurate N-glycan analysis using Rapid™ PNGase F

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This application note describes deglycosylation in minutes using Rapid PNGase F, enabling monitoring of antibody glycosylation, specifically N-glycan profiles.

Read App Note


Transformation of SHuffle® Competent Cell Strains

Last Updated July 2021

Application: Glycobiology and Protein Analysis, Protein Expression and Purification

This application note describes the use of Shuffle Comptetent E.coli Cells to enable the folding of recombinantly expressed proteins in vivo.

Read App Note


Intein-Mediated Protein Ligation IPL and Labeling with the IMPACT™ Kit

Last Updated July 2021

Application: Glycobiology and Protein Analysis, Protein Expression and Purification

This application note provides protocols for intein-mediated protein ligation (IPL) and protein labeling using NEB's IMPACT Kit.

Read App Note


Fully Automated NEBNext® Direct Genotyping Solution using Beckman Coulter® Life Sciences Biomek i7 Workstation

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This application note describes the automation of NEBNext Direct Genotyping Solution using Beckman Coulter Biomek i7 Hybrid workstation for cost-effective, high-throughput, plant and animal applications.

Read App Note


Automated NEBNext® Ultra™ II DNA Library Preparation for Illumina®

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This application note describes an automated NEBNext Ultra II DNA library prep workflow for Illumina, with end repair and dA-tailing steps, on the Freedom EVO NGS workstation.

Read App Note


Automated DNA Library Construction Using the NEBNext Ultra DNA Library Prep Kit for Illumina® on the Beckman Coulter® Biomek FXP Automated Liquid Handler

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note describes automation of DNA library construction on the Beckman Coulter Biomek FXP Dual Arm Multi-Channel 96 and Span-8 automated liquid handler.

Read App Note


Automated Directional RNA-seq Library Preparation using the Biomek FXP Liquid Handler

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note describes the automation of the NEBNext® Ultra Directional RNA Library Prep Kit for Illumina® on the Beckman Coulter® Biomek FXP automated liquid handler.

Read App Note


Improved library quantitation for a broad range of library types using the NEBNext® Library Quant Kit for Illumina®

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note describes the effectiveness of the NEBNext Library Quant Kit for a broad range of library types and sizes.

Read App Note


Monarch® HMW DNA Extraction Kit improves sample preparation for Oxford Nanopore Technologies® sequencing of malaria parasites

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

In this customer-generated technical note, see how the Monarch HMW DNA Extraction Kit was used to extract DNA from Plasmodium and improved nanopore sequencing results for de novo genome assembly.

Read App Note


Improving NGS library performance with lower input amounts using the NEBNext Ultra II RNA Library Prep Kit for Illumina (non-directional)

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note describes non-directional RNA library preparation using poly(A) mRNA enrichment and NEBNext® Ultra II RNA Library Prep Kit for Illumina.

Read App Note


Improved methods for site-directed mutagenesis using Gibson Assembly® Master Mix

Last Updated July 2021

Application: Cloning and Synthetic Biology, NGS Sample Prep and Target Enrichment

This technical note outlines how the Gibson Assembly Master Mix from NEB can reduce time and cost when performing site-directed mutagenesis.

Read App Note


Enhancing Transformation Efficiency

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note provides protocols and tips for enhancing transformation efficiency of competent E. coli cell lines #C2523, C2925, C2984, C2987, C2988, C2992 and C3019.

Read App Note


Nanoliter Scale DNA Assembly Utilizing the NEBuilder® HiFi Cloning Kit with the Labcyte® Echo® 525 Liquid Handler

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes the use of the Labcyte Echo 525 Liquid Handler to miniaturize reaction volumes for two- and five-piece assemblies.

Read App Note


GUIDElines for optimization of Tth Argonaute (TtAgo) reactions

Last Updated July 2021

This technical note provides useful tips to help with designing guides for Tth Argonaute reactions.

Read App Note


Using Glycosidases to Remove, Trim, or Modify Glycans on Therapeutic Proteins

Last Updated July 2021

Application: Glycobiology and Protein Analysis

This application note describes the use of glycosidases on therapeutic proteins to remove glycans for analysis, trim glycans for sequencing, or modify glycans in glycoengineering.

Read App Note


E. coli Lemo21 DE3-A T7 RNA Polymerase-based protein overexpression platform for routine and difficult targets

Last Updated July 2021

Application: Glycobiology and Protein Analysis, Protein Expression and Purification

This application note describes the expression of difficult proteins using the T7 strain, Lemo21(DE3).

Read App Note


Co-expression of Multiple Proteins in Kluyveromyces lactis

Last Updated July 2021

Application: Protein Expression and Purification

This application note describes an easy protocol for the construction of K. lactis strains that co-express multiple heterologous proteins.

Read App Note


Use of the PURExpress® in vitro Protein Synthesis Kit, Disulfide Bond Enhancer and SHuffle® Competent E. coli for heterologous in vitro and in vivo cellulase expression.

Last Updated July 2021

Application: Protein Expression and Purification

This application note describes the use of PURExpress and PDBE to express active proteins in vivo in Shuffle™ competent cells.

Read App Note


A faster workflow for the assessment of genomic loci in mice using a novel HMW DNA extraction technology upstream of Cas9 targeted sequencing

Last Updated July 2021

Application: Genome Editing

This technical note reviews how scientists at The Jackson Laboratory were able to significantly reduce their workflow time for assessment of transgene insert sites in mouse lines using the novel glass-bead based Monarch® HMW DNA Extraction Kit upstream of their Cas9 targeted sequencing workflow.

Read App Note


NEBNext® Enzymatic Methyl-seq (EM-seq™)

Last Updated July 2021

Application: Glycobiology and Protein Analysis, Epigenetics and Cellular Analysis

This technical note describes a new high-performance, enzyme-based methodology developed at NEB that provides an alternative to bisulfite sequencing for methylome analysis.

Read App Note


Biomek i-Series Automated New England Biolabs NEBNext® Ultra™ II DNA Library Prep Kit

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This application note describes the automation of the NEBNext Ultra II DNA Library Prep Kit for the Beckman Biomek i-Series.

Read App Note


Increased transcription detection with the NEBNext® Single Cell/Low Input RNA Library Prep Kit

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note demonstrates use of the NEBNext Single Cell/Low Input RNA Library Prep Kit for high quality library prep from single cells or low inputs.

Read App Note


Bridging dsDNA with a ssDNA Oligo and NEBuilder® HiFi DNA Assembly to create an sgRNA-Cas9 Expression Vector

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes the construction of an sgRNA-Cas9 expression vector using ssDNA oligos and NEBuilder® HiFi DNA Assembly Master Mix.

Read App Note


Improved method for assembly of linear yeast expression cassettes using NEBuilder® HiFi DNA Assembly Master Mix

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes in vitro DNA assembly of expression cassettes using NEBuilder® HiFi DNA Assembly Mix and NEB's K. lactis Protein Expression Kit.

Read App Note


Construction of an sgRNA-Cas9 expression vector via single-stranded DNA oligo bridging of double-stranded DNA fragments

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes an NEB protocol to construct an sgRNA-Cas9 expression vector with ssDNA oligos, a digested vector and NEBuilder® HiFi DNA Assembly Master Mix.

Read App Note


Automated size selection of NEBNext® Small RNA libraries with the Sage Pippin Prep™

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note outlines the use of Pippin Prep beads for size selection of NEBNext Small RNA libraries.

Read App Note


Improved Library Preparation with NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note on DNA library preparation addresses lower input amounts and challenging sample types.

Read App Note


Improving NGS library performance with lower input amounts using the NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (non-directional)

Last Updated July 2021

Application: NGS Sample Prep and Target Enrichment

This technical note describes non-directional RNA library preparation using a rRNA depletion workflow and NEBNext Ultra II RNA Library Prep Kit for Illumina.

Read App Note


Purification of synthetic SARS-CoV-2 viral RNA from biological samples using the Monarch® Total RNA Miniprep Kit and the Monarch RNA Cleanup Kit

Last Updated July 2021

Application: RNA Analysis

This application note describes how Monarch kits can be used for effective viral RNA extraction for SARS-CoV-2 research, including leveraging the QIAcube® and KingFisher™ Flex automation platforms.

Read App Note


Rapid Analysis of Genome Editing Efficiency using PCR Amplicons processed by the Exo-CIP™ Rapid PCR Cleanup Kit followed by Sanger sequencing

Last Updated July 2021

Application: Genome Editing

This application note describes the analysis of genome editing efficiency by using the Exo-CIP™ Rapid PCR Cleanup Kit for PCR cleanup upstream of Sanger sequencing.

Read App Note


SNAPf -based pulse labeling for analysis of protein turnover in living cells

Last Updated July 2021

Application: Epigenetics and Cellular Analysis, DNA Amplification PCR and qPCR, Glycobiology and Protein Analysis

This application note describes the labeling of fusion proteins using an improved version of SNAP-tag, SNAPf.

Read App Note


Using Exonuclease V (RecBCD) to Eliminate Residual Genomic DNA When Purifying Low Copy Plasmids with the Monarch® Plasmid Miniprep Kit

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes the use of Exonuclease V to degrade contaminating gDNA during plasmid miniprep of low copy plasmids.

Read App Note


Breaking through the Limitations of Golden Gate Assembly

Last Updated July 2021

Application: Cloning and Synthetic Biology

Ligase fidelity data and BsaI-HF®v2 provide 20+ fragment Golden Gate assemblies that exhibit high efficiencies, >90% accuracy and low backgrounds.

Read App Note


Efficient Adaptor Ligation for the Preparation of dsDNA Libraries using the Blunt/TA Ligase Master Mix

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes the use of the Blunt/TA Ligase Master Mix for adaptor ligation to dA tailed dsDNA fragments.

Read App Note


Robust Colony PCR from Multiple E. coli Strains using OneTaq® Quick-Load® Master Mixes

Last Updated July 2021

Application: Cloning and Synthetic Biology

This application note describes the use of NEB's OneTaq® Quick-Load® Master Mixes in colony PCR from multiple E. coli strains.

Read App Note


Selective Depletion of Abundant RNAs to Enable Transcriptome Analysis of Low-input and Highly-degraded RNA from FFPE Breast Cancer Samples

Last Updated June 2021

Application: NGS Sample Prep and Target Enrichment

Application note describing selective depletion of abundant RNA from human, mouse and rat, as well as degraded and FFPE breast tumor biopsy RNA samples.

Read App Note


Remove iT PNGase F Effective Release and Recovery of Neutral and Sialylated N glycans

Last Updated June 2021

This application note describes Remove-iT® PNGase F, a new version of PNGase F, that can be easily removed after deglycosylation using chitin magnetic beads.

Read App Note


Proteomics Fast and Efficient Antibody Deglycosylation using Rapid PNGase F

Last Updated June 2021

This application note describes complete removal of N-glycans from antibodies using Rapid™ PNGase F, overcoming lengthy incubation steps needed using PNGase F.

Read App Note


Probe-based qPCR  Probe Compatibility and Multiplexing with Luna Universal Probe qPCR Master Mix

Last Updated June 2021

This application note describes qPCR multiplexing using Luna® Universal Probe qPCR Master Mix for a high performance reaction.

Read App Note


Multiplex PCR using Q5 High-Fidelity DNA Polymerase

Last Updated June 2021

This application note describes the use of NEB's Q5® High-Fidelity DNA Polymerase in multiplex PCR.

Read App Note


Joining of Difficult to Ligate dsDNA Fragments with Blunt TA Ligase Master Mix

Last Updated June 2021

Application: Cloning and Synthetic Biology

This application note describes the use of NEB's Blunt/TA Ligase Master Mix for ligating difficult overhangs and blunt ends.

Read App Note


A Fast One-Step Digestion of DNA or RNA for Global Detection and Characterization of Nucleotide Modifications

Last Updated June 2021

A Fast One-step Digestion of DNA or RNA for Global Detection and Characterization of Nucleotide Modifications

Read App Note


DNA Methylation and Restriction Digests

Last Updated June 2021

Application: Epigenetics and Cellular Analysis

This application note describes how DNA methylation can interfere with restriction digests and how NEB's dam-/dcm- or dam- Competent E.coli cells overcome this.

Read App Note


The Luna 4X RT-qPCR Mix and Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit enable high throughput, automated detection workflows

Last Updated June 2021

This note describes features of the Luna 4X RT-qPCR mix and multiplex assay kit that enable high-throughput, automated SARS-CoV-2 detection workflows.

Read App Note


Quick Ligation Kit

Last Updated June 2021

Application: Cloning and Synthetic Biology

This application note provides protocols for ligation of a variety of cohesive or blunt end DNA using NEB's Quick Ligation™ Kit.

Read App Note


Primer Monitor: an online tool to track SARS-CoV-2 variants that may impact primers used in diagnostic assays

Last Updated June 2021

The Primer Monitor tool can inform SARS-CoV-2 variant studies on primer/probe binding in COVID-19 diagnostic assays, as described in this white paper.

Read App Note


Glycan Analysis of Murine IgG2a by Enzymatic Digestion with PNGase F and Trypsin, Followed by Mass Spectrometric Analysis

Last Updated June 2021

Application: Glycobiology and Protein Analysis

This application note describes the use of PNGase F, trypsin and mass spectroscopy for the release and characterization of glycans on Murine IgG2a.

Read App Note


Glycan Analysis of Murine IgG by Enzymatic Digestion with Endo S and PNGase F Followed by Mass Spectrometric Analysis

Last Updated June 2021

This application note describes the more robust enzymatic removal of glycans on native Murine IgG using Endo S compared to incomplete digestion using PNGase F.

Read App Note


Simultaneous Fluorescent Labeling of Proteins in Living Cells

Last Updated June 2021

This application note describes the simultaneous fluorescent labeling of proteins in living cells using NEB's SNAP-tag® and CLIP-tag™.

Read App Note


Enzymatic PCR cleanup using Exonuclease I and Shrimp Alkaline Phosphatase

Last Updated June 2021

Application: Cloning and Synthetic Biology

This application note describes PCR cleanup using Exonuclease I to degrade residual primers, and Shrimp Alkaline Phosphatase to dephosphorylate remaining dNTPs.

Read App Note


Enzymatic Deglycosylation of a Protein Containing Core 1 O-glycans with O-Glycosidase

Last Updated June 2021

This application note describes the use of a recombinant O-Glycosidase that can cleave core 1 O-linked disaccharides (5) and immature Tn core (GalNAc).

Read App Note


Using the PURExpress Kit

Last Updated June 2021

Application: Protein Expression and Purification

This application note describes the advantages of using PURExpress cell-free in vitro transcription/ translation system for high-throughput screening.

Read App Note


Removal of terminal galactose from a glycoprotein containing tri and tetraantennary N linked sugars with 1 3 6 Galactosidase

Last Updated June 2021

This application note describes the use of an a1-3,6 Galactosidase to remove terminal galactose residues from the tri- and tetra- antennary N-glycoprotein.

Read App Note


Simultaneous Fluorescent Labeling of Proteins in Living Cells

Last Updated June 2021

Application: Epigenetics and Cellular Analysis, DNA Amplification PCR and qPCR, Glycobiology and Protein Analysis

This application note describes the simultaneous fluorescent labeling of proteins in living cells using NEB's SNAP-tag® and CLIP-tag™.

Read App Note


Ph.D.™ Peptide Display Cloning System

Last Updated June 2021

Application: Glycobiology and Protein Analysis

This application note provides protocols for cloning a library of short peptides into M13KE phage using the Ph.D. Peptide Display Cloning System.

Read App Note


Facilitating Detection of SARS-CoV-2 Directly from Patient Samples: Precursor Studies with RT-qPCR and Colorimetric RT-LAMP Reagents

Last Updated May 2021

This paper evaluates the use of NEB RT-qPCR and colorimetric RT-LAMP reagents on unpurified RNA samples through the use of synthetic SARS-Cov-2 viral RNA.

Read App Note


High-throughput qPCR and RT-qPCR Workflows Enabled by Beckman Coulter Echo Acoustic Liquid Handling and NEB Luna Reagents

Last Updated April 2021

This article outlines the use of NEB Luna qPCR and RT-qPCR reagents with the Beckman Coulter Echo Acoustic liquid handler.

Read App Note