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Authors: New England Biolabs, Inc.
Group: New England Biolabs (NEB)
Summary: Cas9 nuclease may be used in vivo to create targeted genome modifications. There are several ways in which to introduce Cas9-guide RNA complexes into cells. Here we present a method for the introduction of Cas9 RNP’s into HEK293 FT cells using the Thermo Fisher Neon Electroporation System. This method uses a final concentration of 20 nM RNP per transfection in a 24-well culture plate.
Proper citation: New England Biolabs, Inc. 2016. Electroporation of Cas9 RNP (ribonucleoprotein) into adherent cells using the Neon® Electroporation. protocols.io https://dx.doi.org/10.17504/protocols.io.frkbm4w Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: The protocol described below will enable degradation of up to 20 pmol of a 25 nt ssDNA (~ 200 ng). In order to degrade larger amounts of ssDNA or ssDNAs longer than 25 nt, we recommend adding more enzyme instead of extending the reaction time. Users should note that ssDNAs longer than 25 nt may form secondary structures that hinder Thermolabile Exonuclease I activity.
Proper citation: New England Biolabs 2020. Protocol for removing ssDNA from dsDNA or RNA Samples (NEB #M0568). protocols.io https://dx.doi.org/10.17504/protocols.io.7ryhm7w Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the "quick" version of Monarch® PCR & DNA Cleanup Kit (5 μg) Protocol (NEB #T1030). For the full protocol, please click here.
Proper citation: New England Biolabs 2020. Quick Protocol for Monarch® PCR & DNA Cleanup Kit (5 ?g) (NEB #T1030). protocols.io https://dx.doi.org/10.17504/protocols.io.bp9emr3e Copy
Authors: Isabel Gautreau
Group: New England Biolabs (NEB)
Summary: Protocol for digestion with NEBNext dsDNA Fragmentase (M0348)
Proper citation: Isabel Gautreau 2015. Digestion with NEBNext dsDNA Fragmentase (M0348). protocols.io https://dx.doi.org/10.17504/protocols.io.cr4v8v Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Ligation protocol for the PCR Cloning Kit (E1202)
Proper citation: New England Biolabs 2015. Ligation Protocol for NEB PCR Cloning Kit (E1202). protocols.io https://dx.doi.org/10.17504/protocols.io.cp5vq5 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the quick version of the Monarch® Plasmid DNA Miniprep Kit Protocol (NEB #T1010). For the full protocol, please click here.
Proper citation: New England Biolabs 2020. Quick Protocol for Monarch® Plasmid Miniprep Kit (NEB #T1010). protocols.io https://dx.doi.org/10.17504/protocols.io.bp9fmr3n Copy
Authors: NEB
Group: New England Biolabs (NEB)
Summary: This is the protocol for DNA Assembly using the NEBuilder® HiFi DNA Assembly Master Mix (E2621).
Proper citation: NEB 2015. NEBuilder HiFi DNA Assembly Reaction (E2621). protocols.io https://dx.doi.org/10.17504/protocols.io.cwaxad Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. T4 PNK Minus Reaction. protocols.io https://dx.doi.org/10.17504/protocols.io.cp3vqm Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: The following is a "typical" restriction endonuclease reaction. Please see the "guidelines" tab below for the NEB tips on optimizing restriction digests.
Proper citation: New England Biolabs 2014. Restriction Digest. protocols.io https://dx.doi.org/10.17504/protocols.io.chkt4v Copy
Authors: Isabel Gautreau
Group: New England Biolabs (NEB)
Summary: Protocol for digestion with NEBNext dsDNA Fragmentase (M0348)
Proper citation: Isabel Gautreau 2016. Digestion with NEBNext dsDNA Fragmentase (M0348). protocols.io https://dx.doi.org/10.17504/protocols.io.e9rbh56 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: The NEBNext Ultra II Directional RNA Library Prep Kit for Illumina contains the enzymes and buffers required to convert a broad range of input amounts of RNA into high quality directional (strand-specific) libraries for next-generation sequencing on the Illumina platform. The fast, user-friendly workflow has minimal hands-on time and is compatible with poly(A) mRNA enrichment and rRNA depletion methods.
Proper citation: New England Biolabs 2018. Protocol for use with rRNA Depleted FFPE RNA and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760, E7765). protocols.io https://dx.doi.org/10.17504/protocols.io.ubaesie Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the PCR protocol for Phusion® High-Fidelity DNA Polymerase (M0530)
Proper citation: New England Biolabs 2015. PCR with Phusion® Polymerase (M0530). protocols.io https://dx.doi.org/10.17504/protocols.io.cdjs4m Copy
Authors: Isabel Gautreau
Group: New England Biolabs (NEB)
Proper citation: Isabel Gautreau 2020. NEBNext Ultra End Prep Mixture E7442. protocols.io https://dx.doi.org/10.17504/protocols.io.bpiumkew Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: This is the protocol for the Q5® Site-Directed Mutagenesis Kit (E0554)
Proper citation: New England Biolabs 2014. Q5® Site-Directed Mutagenesis (E0554). protocols.io https://dx.doi.org/10.17504/protocols.io.cjpumm Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. OneTaq Mixture M0480. protocols.io https://dx.doi.org/10.17504/protocols.io.cq9vz5 Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. NEB Immunoprecip. Buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.crbv2m Copy
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. NEBNext rRNA Depletion Kit (Human/Mouse/Rat) E6310. protocols.io https://dx.doi.org/10.17504/protocols.io.mzkc74w Copy
Authors: NEB
Group: New England Biolabs (NEB)
Summary: This is the synthesis protocol for modified nucleotides using the HiScribe™ T7 Quick High Yield RNA Synthesis Kit (E2050). The kit is capable of synthesizing biotin- or dye-modified RNA.
Proper citation: NEB 2015. RNA Synthesis with Modified Nucleotides (E2050). protocols.io https://dx.doi.org/10.17504/protocols.io.cv7w9m Copy
Authors: Lauren Moran, 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: Lauren Moran, New England Biolabs 2018. Protocol for use with NEBNext rRNA Depletion Kit (Human/Mouse/Rat) E6310 (E7760). protocols.io https://dx.doi.org/10.17504/protocols.io.sxuefnw Copy
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Proper citation: New England Biolabs 2015. T4 PNK Radioactive Reaction. protocols.io https://dx.doi.org/10.17504/protocols.io.cp8vrv Copy
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