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Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: PCR Protocol for Taq DNA Polymerase with ThermoPol® Buffer (M0267)
Proper citation: New England Biolabs 2015. PCR with Taq and ThermoPol® Buffer (M0267). protocols.io https://dx.doi.org/10.17504/protocols.io.cifubm Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2020. NEB Monarch® RNA Cleanup Kit. protocols.io https:// Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: The NEBNext Ultra II FS DNA Library Prep Kit for Illumina contains the enzymes and buffers required to convert a broad range of input amounts of DNA into high quality libraries for next-generation sequencing on the Illumina platform. The fast, user-friendly workflow also has minimal hands-on time.Note: The Ultra II FS Kit is not compatible with bisulfite conversion workflows.
Proper citation: New England Biolabs 2021. E7805 NEBNext® Ultra™ II FS DNA Library Prep Kit for Illumina® Protocol to use with Inputs ≥ 100 ng (Chapter 2). protocols.io https://dx.doi.org/10.17504/protocols.io.nwxdffn Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Rapid and irreversible heat inactivation eliminates unwanted activityImproved storage stability versus native enzymeFaster reaction setup (no supplemental additives like zinc required) and shorter incubation timeFlexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)Less enzyme required (high specific activity), resulting in a lower cost per reactionNo need for multiple phosphatases (Quick CIP removes 5′- and 3′- phosphates from DNA, RNA and dNTPs )Active on unincorporated dNTPs in PCR products - improves DNA sequencing and SNP analysisRecombinant for purity, consistency and value
Proper citation: New England Biolabs 2020. Enzymatic PCR Cleanup Protocol (NEB #M0525). protocols.io https://dx.doi.org/10.17504/protocols.io.7r4hm8w Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Protocol for Dephosphorylation of 5´-ends of DNA using CIP in Restriction Enzyme Reaction. Uses the Calf Intestinal Alkaline Phosphatase (CIP - M0290)
Proper citation: New England Biolabs 2014. Dephosphorylation using CIP in Restriction Enzyme Reaction (M0290). protocols.io https://dx.doi.org/10.17504/protocols.io.cdhs35 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the protocol for dephosphorylation of 5'-ends of DNA using AnP (Antarctic Phosphatase - M0289).
Proper citation: New England Biolabs 2014. Dephosphorylation using AnP (M0289). protocols.io https://dx.doi.org/10.17504/protocols.io.cdns5d Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Both the MBP-tag and TEV Protease are polyhistidine-tagged for easy removal from the TEV Protease reaction. Loading the digest onto NEBExpress Ni Resin (NEB #S1428) sequesters both the MBP-tag and TEV Protease, thereby isolating the protein of interest in the column flow through.
The binding capacity of NEBExpress Ni Resin is ≥ 10 mg/ml. The binding capacity will vary depending on the target protein, binding conditions and the accessibility of the His-tag.
It is recommended to estimate the amount of the His-tagged MBP released by TEV Protease cleavage by first analyzing a sample by SDS-PAGE and comparing its band size and intensity to that of the provided MBP6 protein control.
Proper citation: New England Biolabs 2020. Separating the Protein of Interest from MBP after TEV Protease Cleavage (NEB #E8201). protocols.io https://dx.doi.org/10.17504/protocols.io.bfebjjan Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2020. Protocol for T7 Exonuclease (NEB #M0263). protocols.io https:// Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Protocol for non-radioactive phosphorylation with T4 PNK 3' phosphatase minus.
Proper citation: New England Biolabs 2015. Non-radioactive phosphorylation with T4 PNK 3' phosphatase minus. protocols.io https://dx.doi.org/10.17504/protocols.io.cp2vqd 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 https://dx.doi.org/10.17504/protocols.io.bfa4jigw Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. E1202 Blunting Mixture. protocols.io https://dx.doi.org/10.17504/protocols.io.crqv5v 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 https://dx.doi.org/10.17504/protocols.io.bsbrnam6 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. Mixture for PCR Amplification (E7370). protocols.io https://dx.doi.org/10.17504/protocols.io.cg7tzm Copy
Authors: New England Biolabs
Proper citation: New England Biolabs 2020. Protein Synthesis Reaction using PURExpress (E6800). protocols.io https:// Copy
Authors: New England Biolabs
Proper citation: New England Biolabs 2020. NEBExpress Ni Spin Column Reaction Protocol (NEB #S1427). protocols.io https:// 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 https://dx.doi.org/10.17504/protocols.io.7sehnbe Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: OneTaq DNA Polymerase allows for greater amplification sensitivity across a wide variety of amplicons regardless of GC content.
Proper citation: New England Biolabs 2015. PCR Protocol for OneTaq® DNA Polymerase (M0480). protocols.io https://dx.doi.org/10.17504/protocols.io.cq8vzv Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Protocol for blunting ends by 3' overhang removal and fill-in of 3' recessed (5' overhang) ends using DNA Polymerase I, Large (Klenow) Fragment (M0210)
Proper citation: New England Biolabs 2015. Large (Klenow) Fragment Blunting (M0210). protocols.io https://dx.doi.org/10.17504/protocols.io.cpgvjv Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Quick Protocol for Oligonucleotide Cleanup Using the Monarch® PCR & DNA Cleanup Kit (5 μg) (NEB #T1030)
Proper citation: New England Biolabs 2020. Quick Protocol for Oligonucleotide Cleanup Using the Monarch® PCR & DNA Cleanup Kit (5 ?g) (NEB #T1030). protocols.io https://dx.doi.org/10.17504/protocols.io.bp9dmr26 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 https://dx.doi.org/10.17504/protocols.io.crtv6m Copy
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