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Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: The NEBNext rRNA Depletion Kit (Human/Mouse/Rat) depletes both cytoplasmic (5S rRNA, 5.8S rRNA, 18S rRNA and 28S rRNA) and mitochodrial ribosomal RNA (12S rRNA and 16S rRNA) from human, mouse and rat total RNA preparations. This product is suitable for both intact and degraded RNA (e.g. FFPE RNA). The resulting rRNA-depleted RNA is suitable for RNA-Seq, random-primed cDNA synthesis, or other downstream RNA analysis applications.
Proper citation: New England Biolabs 2018. Protocol for use with NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (E6310) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760, E7765). protocols.io dx.doi.org/10.17504/protocols.io.wkdfcs6 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. T4 PNK Minus Reaction. protocols.io dx.doi.org/10.17504/protocols.io.cp3vqm Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: The NEBNext Multiplex Small RNA Library Prep Set for Illumina contains the adaptors, primers, enzymes and buffers required to convert small RNAs into indexed libraries for next generation sequencing on the Illumina platform. The novel workflow has been optimized to minimized adaptor dimers, while producing high-yield, high-diversity libraries. The unique workflow of the NEBNext® Small RNA library prep kits addresses the challenge of minimization of adaptor-dimers while achieving production of high-yield, diverse multiplex libraries in a simple protocol. Minimized adaptor-dimer contaminationHigh yieldsInput RNA can be Total RNASuitable for methylated small RNAs (e.g., piRNAs) as well as unmethylated small RNAsMultiplex Small RNA Library Prep Workflow This kit includes a novel protocol that results in higher yields and lower adaptor-dimer contamination.
Proper citation: New England Biolabs 2018. Protocol for use with NEBNext® Small RNA Library Prep Set for Illumina® (E7300, E7580, E7560, E7330). protocols.io dx.doi.org/10.17504/protocols.io.t7qermw Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2014. Cas9 M0386 Mixture. protocols.io dx.doi.org/10.17504/protocols.io.ch3t8m Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Please note that protocols with Q5U Hot Start High-Fidelity DNA Polymerase may differ from protocols with other polymerases. Conditions recommended in the protocols should be used for optimal performance.
Proper citation: New England Biolabs 2020. PCR Using Q5U Hot Start High-Fidelity DNA Polymerase (NEB #M0515). protocols.io dx.doi.org/10.17504/protocols.io.7sehnbe Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the correct protocol if you are using the C2987H cells. If you are using the C2987I cells, please refer to this protocol.
Proper citation: New England Biolabs 2014. High Efficiency Transformation Protocol (C2987H). protocols.io dx.doi.org/10.17504/protocols.io.chgt3v Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: TEV Protease, also known as Tobacco Etch Virus (TEV) Protease, is a highly specific cysteine protease that recognizes the amino-acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) and cleaves between the Gln and Gly/Ser residues. It is often used for the removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins. TEV Protease has a 7xHis-tag for easy removal from a reaction using nickel affinity resins and has been engineered to improve thermal stability and decrease autolysis.
Proper citation: New England Biolabs 2020. Cleavage of the Fusion Protein (TEV Protease). protocols.io dx.doi.org/10.17504/protocols.io.bfd9ji96 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the protocol for making two 250 ml cultures of electrocompetent cells
Proper citation: New England Biolabs 2015. Making your own electrocompetent cells. protocols.io dx.doi.org/10.17504/protocols.io.crtv6m Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the Double Digest Protocol with Standard Restriction Enzymes, in the same buffer, using two separate incubation temperatures for the enzymes.
Proper citation: New England Biolabs 2014. Multi-temperature Double Digest (single buffer). protocols.io dx.doi.org/10.17504/protocols.io.cjkukv Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: NEBExpress® Ni Resin is an affinity matrix for the isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins. It is intended for use in gravity or pressure flow columns, and batch purifications. NEBExpress Ni Resin is comprised of a highly uniform and stable chemical-tolerant resin, pre-charged with nickel ions on the matrix surface. It is resistant to a wide range of chemicals, including NaOH, EDTA, DTT and β-Mercaptoethanol.Purification of ≥10 mg His-tagged protein per 1 ml of resinIntended for use in gravity or pressure flow columns, and batch purificationsHigh 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 reagentsIsolation and purification of His-tagged fusion proteins under native or denaturing conditions
Proper citation: New England Biolabs 2020. NEBExpress Ni Resin Pressurized Column Typical Protocol (NEB #S1428). protocols.io dx.doi.org/10.17504/protocols.io.bfa4jigw Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. BioBrick E0546 Downstream Reaction. protocols.io dx.doi.org/10.17504/protocols.io.cpkvkv Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB), Coronavirus Method Development Community
Summary: This protocol details methods for the NEBNext® ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies®), NEB #E7660S/L 24/96 reactions.
Proper citation: New England Biolabs 2021. NEBNext® ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies®) E7760. protocols.io dx.doi.org/10.17504/protocols.io.bsbrnam6 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the Double Digest Protocol with Standard Restriction Enzymes, using a common reaction and same incubation temperature for both enzymes.
Proper citation: New England Biolabs 2014. Single-temperature Double Digest. protocols.io dx.doi.org/10.17504/protocols.io.cjhuj5 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Quick Protocol for Monarch® Total RNA Miniprep Kit (NEB #T2010).Quickly and easily purify up to 100 µg of high-quality total RNA from multiple sample types – all with one kit!For use with blood, cells and tissuesAlso works with tough to lyse samples (bacteria, yeast, plant)Effectively purifies total RNA of all sizes, including small RNAs >20 ntEfficient genomic DNA removal (column and DNase I-based)Contains Proteinase K for processing of tissues and blood samplesIncludes RNA Protection Reagent for sample preservationExcellent valueKit components available separately
Proper citation: New England Biolabs 2018. Quick Protocol for Monarch® Total RNA Miniprep Kit (NEB #T2010). protocols.io dx.doi.org/10.17504/protocols.io.nyadfse Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: T7 Endonuclease I recognizes and cleaves non-perfectly matched DNA. This protocol describes how to determine genome targeting efficiency by digesting annealed PCR products with T7 Endonuclease I. In the first step PCR products are produced from the genomic DNA of cells whose genomes were targeted using Cas9, TALEN, ZFN etc. In the second step, the PCR products are annealed and digested with T7 Endonuclease I. Fragments are analyzed to determine the efficiency of genome targeting.
Proper citation: New England Biolabs 2016. Determining Genome Targeting Efficiency using T7 Endonuclease I (M0302). protocols.io dx.doi.org/10.17504/protocols.io.etbbein Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. Mixture for PCR Amplification (E7370). protocols.io dx.doi.org/10.17504/protocols.io.cg7tzm Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the synthesis protocol using the HiScribe™ T7 Quick High Yield RNA Synthesis Kit (E2050)
Proper citation: New England Biolabs 2015. Standard RNA Synthesis (E2050). protocols.io dx.doi.org/10.17504/protocols.io.cr2v8d Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This protocol is for First Strand Synthesis with Reverse Transcriptase
Proper citation: New England Biolabs 2015. First Strand Synthesis with Reverse Transcriptase. protocols.io dx.doi.org/10.17504/protocols.io.cjuunv Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. Loading Marker Mixture. protocols.io dx.doi.org/10.17504/protocols.io.cq5vy5 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Quick Protocol for Monarch® Total RNA Miniprep Kit (NEB #T2010).Quickly and easily purify up to 100 µg of high-quality total RNA from multiple sample types – all with one kit!For use with blood, cells and tissuesAlso works with tough to lyse samples (bacteria, yeast, plant)Effectively purifies total RNA of all sizes, including small RNAs >20 ntEfficient genomic DNA removal (column and DNase I-based)Contains Proteinase K for processing of tissues and blood samplesIncludes RNA Protection Reagent for sample preservationExcellent valueKit components available separately
Proper citation: New England Biolabs 2020. Quick Protocol for Monarch® Total RNA Miniprep Kit (NEB #T2010). protocols.io dx.doi.org/10.17504/protocols.io.bp9cmr2w Copy
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