The abundance of ribosomal RNAs (rRNAs), constituting 80–90% of total RNA, necessitates removal upstream of next generation sequencing and other applications. This second generation rRNA depletion kit incorporates reagent, probe and protocol improvements, resulting in superior depletion performance. The efficient RNAse-H-based workflow, and close spacing of probes, enables effective depletion from both low- and high-quality RNA, with a broad range of input amounts. 

For added convenience, this kit contains NEBNext RNA Sample Purification Beads (Agencourt^® RNAClean^® XP beads from Beckman Coulter).

• Superior depletion of rRNA from human, mouse and rat RNA
• Depletion of cytoplasmic (5S, 5.8S, 18S, 28S, human ITS, ETS) and mitochondrial (12S and 16S) rRNA
• Compatible with a broad range of input amounts: 10 ng - 1 µg
• Suitable for low-quality or high-quality RNA
• Fast workflow: 2 hours, with less than 10 minutes hands-on time

The kit is also available without RNAClean^® beads. 

Ribosomal RNA (rRNA) is highly abundant in bacterial samples, and its removal is desirable in order to reveal the biological significance of less abundant transcripts. Specific enrichment of bacterial mRNAs is challenging due to their lack of poly(A) tails, and so the converse - efficient and specific removal of bacterial rRNA – is necessary.

The NEBNext^® rRNA Depletion Kit (Bacteria) employs the NEBNext RNase H-based RNA depletion workflow to target removal of rRNA from gram-positive and gram-negative organisms. The method is effective with intact and degraded RNA, from monocultures or samples with mixed bacterial species (e.g., metatranscriptomic).

• Efficient, specific depletion of bacterial rRNA (5S, 16S, 23S)
• Compatible with both gram-positive and gram-negative organisms
• Effective with monocultures and mix of bacterial species (e.g., metatranscriptome)
• Compatible with a broad range of input amounts: 10 ng - 1 µg
• Suitable for low-quality or high-quality RNA
• Fast workflow: 2 hours, with less than 10 minutes hands-on time

The kit is also available with RNAClean^® beads.

Ribosomal RNA (rRNA) is highly abundant in bacterial samples, and its removal is desirable in order to reveal the biological significance of less abundant transcripts. Specific enrichment of bacterial mRNAs is challenging due to their lack of poly(A) tails, and so the converse - efficient and specific removal of bacterial rRNA – is necessary.

The NEBNext^® rRNA Depletion Kit (Bacteria) employs the NEBNext RNase H-based RNA depletion workflow to target removal of rRNA from gram-positive and gram-negative organisms. The method is effective with intact and degraded RNA, from monocultures or samples with mixed bacterial species (e.g., metatranscriptomic).

For added convenience, this kit contains NEBNext RNA Sample Purification Beads (Agencourt® RNAClean® XP beads from Beckman Coulter).

• Efficient, specific depletion of bacterial rRNA (5S, 16S, 23S)
• Compatible with both gram-positive and gram-negative organisms
• Effective with monocultures and mix of bacterial species (e.g., metatranscriptome)
• Compatible with a broad range of input amounts: 10 ng - 1 µg
• Suitable for low-quality or high-quality RNA
• Fast workflow: 2 hours, with less than 10 minutes hands-on time

The kit is also available without RNAClean^® XP beads.

The NEBExpress® Cell-free E. coli Protein Synthesis System is an extract-based transcription/translation system derived from E. coli cells that have been engineered for high in vitro synthesis performance. It is designed to synthesize proteins ranging in size from 17-230 kDa in just 2-4 hours under the control of T7 RNA Polymerase.

• Synthesize high yields of protein (typically 0.5 mg/ml) 
• Protein can be synthesized and visualized in approximately 2 - 4 hours
• Synthesize various target proteins ranging from 17 to 230 kDa
• Templates can be plasmid DNA, linear DNA or mRNA.
• Minimal protease activity ensures stability of desired target protein
• RNase contamination can be inhibited by the supplied RNase inhibitor, eliminating clean-up steps
• Protein can be isolated with affinity purification or subject to direct functional analysis
• Reactions can be directly loaded onto an SDS-PAGE gel without the need for acetone or TCA precipitation
• Flexible reaction conditions achieve maximum yield; protein synthesis can be sustained for 10 hours at 37°C or up to 24 hours at lower temperatures
• Reactions can be miniaturized or scaled up
• Compatible with the PURExpress Disulfide Bond Enhancer (NEB #E6820) and NEBExpress GamS Nuclease Inhibitor (NEB #P0774)

Catalog #E9530

NEBNext Direct^® target enrichment is now available for targeted genotyping!

The NEBNext Direct Genotyping Solution delivers cost-effective, high-throughput, sequence-based target genotyping for a wide variety of applications in plant, animal, and human.

• Ideal solution for genotyping hundreds to thousands of markers
• Reduce costs and streamline workflow through pre-capture pooling of up to 96 samples
• Maximize sequencer efficiency through dual barcode sample indexing plus Unique Molecular Identifier
• Unparalleled target coverage uniformity through unique capture-based enrichment
• Eliminate marker dropouts with finely tuned bait design
• Increase sample throughput using 1-day, automatable workflow

Interested in evaluating the NEBNext Direct Genotyping Solution?

Contact Us

The NEBExpress^® MBP Fusion and Purification System takes advantage of the strong Ptac promoter and the translation initiation signals of maltose binding protein (MBP) to enhance solubility and expression levels of a desired protein in E. coli. The resulting product is an MBP fusion protein, which is then purified by affinity chromatography.

• Reliable E. coli expression: substantial yields (up to 100 mg/L)
• Fusion to MBP has been shown to enhance the solubility of proteins expressed in E. coli ⁽¹⁾
• Two-step purification: amylose elution followed by TEV Protease cleavage and Ni resin isolation results in a highly pure tag-free target protein
• Gentle elution with maltose; no detergents or harsh denaturants required

• Coordinated cell lysis, RNA release, and genomic DNA removal in a fast 15-minute protocol
• Effective cell lysis preparation from 10 to 100,000 cells across numerous cell lines
• Minimal sample loss compared to alternative RNA extraction methods, especially at low cell input
• Compatible with high-throughput applications: cell lysis can take place at room temperature
• Optimal performance when paired with Luna Universal One-Step RT-qPCR kits: NEB #E3005 (dye-based) or NEB #E3006 (probe-based)
• Complete lysis and one-step RT-qPCR workflow available: NEB #E3030 (dye-based) or NEB #E3031 (probe-based) 

Go direct from cells to RNA quantitation without purification

• Contains reagents for 100 lysis preps and 500 one-step RT-qPCR reactions (probe)
• Effective cell lysis preparation from 10 to 100,000 cells across numerous cell lines
• Coordinated cell lysis, RNA release, and genomic DNA removal in a fast 15-minute protocol
• Features Luna WarmStart^® RT paired with Hot Start Taq for increased thermostability and room temperature setup
• Additional lysis preps available: Luna Cell Ready Lysis Module (NEB #E3032)

Go direct from cells to RNA quantitation without purification

• Contains reagents for 100 lysis preps and 500 one-step RT-qPCR reactions (dye)
• Effective cell lysis preparation from 10 to 100,000 cells across numerous cell lines
• Coordinated cell lysis, RNA release, and genomic DNA removal in a fast 15-minute protocol
• Features Luna WarmStart^® RT paired with Hot Start Taq for increased thermostability and room temperature setup
• Additional lysis preps available: Luna Cell Ready Lysis Module (NEB #E3032)

Learn about ligase fidelity and push the limits of your Golden Gate Assembly Reactions (20+ fragments)

• Uses BsmBI-v2, which is optimized for Golden Gate Assembly
• Use for seamless cloning – no scar remains following assembly
• Ideal for ordered assembly of multiple fragments (2-20+) in a single reaction
• Can also be used for cloning of single inserts and library preparations
• Efficient with regions with high GC content and areas of repeats
• Compatible with a broad range of fragment sizes (< 100 bp to > 15 kb)
• Includes destination plasmid compatible with a variety of Type IIS restriction enzymes commonly used in Golden Gate Assembly
• Free tool available at GoldenGate.neb.com

Try our free Ligase Fidelity Tools to assist with design of high fidelity Golden Gate Assemblies

Rapid, sensitive and precise probe-based qPCR detection and quantitation of target RNA targets. 

Make a simpler choice

• For instruments that do not require the ROX passive reference dye
• Convenient master mix formats and user-friendly protocols simplify reaction setup
• Non-interfering, visible tracking dye helps to eliminate pipetting errors

Experience best-in-class performance

• All Luna^® products have undergone rigorous testing to optimize specificity, sensitivity, accuracy and reproducibility
• Products perform consistently across a wide variety of sample sources
• A comprehensive evaluation of commercially-available qPCR and RT-qPCR reagents demonstrates superior performance of Luna products

Optimize your RT-qPCR with Luna WarmStart^® Reverse Transcriptase

• Novel, thermostable reverse transcriptase (RT) improves performance 
• WarmStart RT paired with Hot Start Taq increases reaction specificity and robustness

The abundance of ribosomal RNAs (rRNAs), constituting 80–90% of total RNA, necessitates removal upstream of next generation sequencing and other applications. This second generation rRNA depletion kit incorporates reagent, probe and protocol improvements, resulting in superior depletion performance. The efficient RNAse-H-based workflow, and close spacing of probes, enables effective depletion from both low- and high-quality RNA, with a broad range of input amounts. 

• Superior depletion of rRNA from human, mouse and rat RNA
• Depletion of cytoplasmic (5S, 5.8S, 18S, 28S, human ITS, ETS) and mitochondrial (12S and 16S) rRNA
• Compatible with a broad range of input amounts: 10 ng - 1 µg
• Suitable for low-quality or high-quality RNA
• Fast workflow: 2 hours, with less than 10 minutes hands-on time

The kit is also available with RNAClean^® beads. 

For optimal workflow flexibility, NEBNext^® Multiplex Oligos are available separately from library preparation kits, providing adaptors and primers for high-yield multiplex Illumina^® library production. The NEBNext Adaptor has a unique hairpin loop structure that minimizes adaptor-dimer formation, while NEBNext index PCR primers enable index (barcode) incorporation during library amplification. This third set of unique-dual indices (UDIs) includes 96 pre-mixed unique pairs of index primers (i5 and i7), packaged in a single-use 96-well plate with a pierceable foil seal. 

Additional NEBNext Multiplex Oligos products are available in sets of 12 and 96 single index primers as well as additional dual index primers for increased sample identification specificity.

• Enables complete filtering of index-hopped reads
• Highly efficient adaptor ligation
• Minimized adaptor-dimer formation
• Includes index primers for library multiplexing
• Combine with sets 1, 2, and 4 for 384 barcodes

For optimal workflow flexibility, NEBNext^® Multiplex Oligos are available separately from library preparation kits, providing adaptors and primers for high-yield multiplex Illumina^® library production. The NEBNext Adaptor has a unique hairpin loop structure that minimizes adaptor-dimer formation, while NEBNext index PCR primers enable index (barcode) incorporation during library amplification.  This fourth set of unique-dual indices (UDIs) includes 96 pre-mixed unique pairs of index primers (i5 and i7), packaged in a single-use 96-well plate with a pierceable foil seal. 

Additional NEBNext Multiplex Oligos products are available in sets of 12 and 96 single index primers as well as additional dual index primers for increased sample identification specificity.

• Enables complete filtering of index-hopped reads
• Highly efficient adaptor ligation
• Minimized adaptor-dimer formation
• Includes index primers for library multiplexing
• Combine with sets 1, 2, and 3 for 384 barcodes

For optimal workflow flexibility, NEBNext^® Multiplex Oligos are available separately from library preparation kits, providing adaptors and primers for high-yield multiplex Illumina^® library production. The NEBNext Adaptor has a unique hairpin loop structure that minimizes adaptor-dimer formation, while NEBNext index PCR primers enable index (barcode) incorporation during library amplification. This second set of unique-dual indices (UDIs) includes 96 pre-mixed unique pairs of index primers (i5 and i7), packaged in a single-use 96-well plate with a pierceable foil seal. 

Additional NEBNext Multiplex Oligos products are available in sets of 12 and 96 single index primers as well as additional dual index primers.

• Increased sample identification specificity
• Enables complete filtering of index-hopped reads
• Highly efficient adaptor ligation
• Minimized adaptor-dimer formation
• Includes index primers for library multiplexing

Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality.

• Time-Saver™ qualified for digestion in 5-15 minutes
• Used in Golden Gate Assembly
• Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
• Supplied with 1 vial of Gel Loading Dye, Purple (6X) (NEB #B7024)

• Purification of ≥10 mg His-tagged protein per 1 ml of resin
• Intended for use in gravity or pressure flow columns, and batch purifications
• High specific binding of His-tagged proteins yielding purities of >95%
• Strong nickel ion binding provides excellent resistance to EDTA and reducing agents. Compatible with commercially available detergent-based cell lysis reagents
• Isolation and purification of His-tagged fusion proteins under native or denaturing conditions

• Includes ready-to-use pre-packed Ni spin columns and all required buffers for purification
• Purification of ≥1 mg His-tagged protein per column in as little as 15 minutes
• High specific binding of His-tagged proteins yielding purities of >95%
• Strong nickel ion binding provides excellent resistance to EDTA and reducing agents. Compatible with commercially available detergent-based cell lysis reagents
• Isolation and purification of His-tagged fusion proteins under native or denaturing conditions

Hi-T7 RNA Polymerase is designed for in vitro transcription of RNA at higher temperatures.

• Active from 37-56°C, optimal incubation temperature is 50-52°C.
• In vitro transcription using T7 phage promoter
• High concentration version of Hi-T7 RNA Polymerase (NEB #M0658)
• Increased co-transcriptional capping efficiency with cap analogs
• Decreased immunogenicity from RNA synthesized at higher temperature due to reduced dsRNA by-product formation
• Recommended for experienced users with knowledge of transcription optimization 
• Hi-T7 RNA Polymerase Reaction buffer and magnesium chloride included

• Salt-T4 DNA Ligase is an engineered variant of T4 DNA Ligase with improved salt tolerance 
• Capable of joining blunt end and cohesive end termini, as well as repairing single stranded nicks in duplex DNA and some DNA/RNA hybrids (1).
• Salt-T4 DNA Ligase seals nicks for these DNA substrates.
• Supplied with both T4 DNA Ligase Buffer and StickTogether™ DNA Ligase Buffer.
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart.

• StickTogether™ Buffer is supplied with T3 (NEB #M0317), T7 (NEB #M0318), Hi-T4™ (NEB #M2622) and Salt-T4^® (NEB #M0467) DNA Ligases    
• At a 1X concentration, this reaction buffer ensures optimal activity of these DNA ligases
• Provided as a 2X solution
• Contains 2 vials of buffer

In vitro transcription using T7 phage promoter

• High concentration version of T7 RNA Polymerase (NEB #M0251)
• Recommended for experienced users with knowledge of transcription optimization. 
• RNA Polymerase Reaction Buffer and magnesium chloride included

NEBExpress GamS Nuclease Inhibitor is a recombinant protein that inhibits Exonuclease V (RecBCD) activity and stabilizes linear DNA templates in E. coli based in vitro protein synthesis reactions.  

• Enhances synthesis yield of linear DNA using the NEBExpress Cell-free E. coli Protein Synthesis System
• Recombinant enzyme; ≥ 95% purity, as determined by SDS-PAGE 
• No detectable protease activity, ensuring stability of desired target protein
• Compatible with various target proteins, ranging from 17 to 230 kDa
• Optimal storage buffer for performance in protein synthesis reactions
• Flexible reaction conditions achieve maximum yield; activity can be sustained for 10 hours at 37°C or up to 24 hours at lower temperatures

• ≥ 99% purity as determined by HPLC analysis
• For use with polymerases such as Taq DNA Polymerase (NEB# M0273) or Q5U^® Hot Start High-Fidelity DNA Polymerase (NEB# M0515)
• Combine with Antarctic Thermolabile UDG (NEB# M0372) to prevent carryover contamination in PCR and qPCR-related technologies

The pMAL-c6T Vector is designed to produce maltose-binding protein (MBP) fusions in the cytoplasm. The MBP has been engineered for tighter binding to amylose resin. The vector expresses an N-terminal his-tagged malE gene followed by a multiple cloning site containing a TEV protease recognition sequence, allowing MBP to be cleaved from the protein of interest after purification. 

• pMAL-c6T is the recommended vector for use with the NEBExpress^® MBP Fusion and Purification System (NEB #E8201)
• Fusion to MBP significantly enhances the solubility and proper folding of proteins expressed in E. coli ⁽¹⁾
• Vector includes His-tag and TEV Protease recognition site, allowing the protein of interest to be cleaved from MBP using TEV Protease (NEB #P8112)
• Two-step purification: amylose resin elution followed by TEV Protease cleavage and Ni resin isolation results in a highly-pure, tag-free target protein
• MBP has been engineered for tighter binding to amylose resin
• Ampicillin resistant

• Decaps RNA that contains ADP-containing, non-canonical initiating nucleotides (including NAD⁺/NADH caps), resulting in ligatible monophosphate at the 5′ end.
• Hydrolyzes NADH and NAD⁺ into AMP and reduced nicotinamide monophosphate nucleoside (NMNH), and AMP and nicotinamide monophosphate nucleoside (NMN⁺), respectively.
• Hydrolyzes diphosphate dinucleotides and metabolic co-factors containing an ADP moiety such as AppA, FAD, Coenzyme A, etc.
• Useful for NAD⁺ decapping or deNADding when NAD⁺ is the 5′ initiating nucleotide of RNA.
• Useful for detection and identification of RNA containing non-canonical initiating nucleotides using ligation-based methods such as 5′ RACE and RNA-seq.

This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.

Thermus thermophilus argonaute (TtAgo) is a programmable DNA-endonuclease which requires a short 5’-phosphorylated single-stranded DNA guide to target its activity to a specific corresponding sequence on a substrate. TtAgo introduces one break in the phosphodiester backbone of the complementary substrate sequence.

• Short 16–18 nucleotide 5′-phosphorylated ssDNA guides are cost effective and can be phosphorylated with NEB T4 Polynucleotide Kinase
• Guide/target sequence selection is not limited by the requirement of an adjacent sequence motif
• Highly active on ssDNA and most dsDNA substrates (generates a nick in dsDNA substrates), with mild activity on ssRNA substrates
• Recommended for in vitro applications
• Refer to GUIDElines for optimization of Tth Argonaute (TtAgo) reactions

• Hi-T4 DNA Ligase is an engineered variant of T4 DNA Ligase with improved thermostability compared to T4 DNA Ligase
• Capable of joining blunt end and cohesive end termini, as well as repairing single stranded nicks in duplex DNA and some DNA/RNA hybrids (1)
• Hi-T4 DNA Ligase seals nicks for these DNA substrates
• Supplied with both T4 DNA Ligase Buffer and StickTogether™ DNA Ligase Buffer
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart