Programmable Nucleases
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FAQs for
Programmable Nucleases
- What are the differences between EnGen® Lba Cas12a (Cpf1) (NEB #M0653) and EnGen Spy Cas9 NLS (NEB #M0646)?
- Which nuclear localization signal is fused to EnGen® Lba Cas12a (Cpf1)?
- Why am I observing limited or no digestion with EnGen® Lba Cas12a (Cpf1)?
- Why does digestion/editing efficiency differ between two different guide RNAs?
- Can mutations generated with EnGen Lba Cas12a (Cpf1) be detected using T7 Endonuclease I (NEB #M0302) or the EnGen Mutation Detection Kit (NEB #E3321S)?
- Can gRNA for use with EnGen Lba Cas12a (Cpf1) be generated using the EnGen sgRNA Synthesis Kit, S. pyogenes (NEB #E3322)?
- How do I design a guide RNA for use with EnGen Lba Cas12a?
- How do I dilute the enzyme to 1 μM for in vitro reactions?
- Can the EnGen® sgRNA Synthesis Kit (NEB #E3322S) be used to synthesize sgRNAs specific for Sau Cas9?
- How can I synthesize a sequence-specific sgRNA for experiments using EnGen® Sau Cas9?
- How can the genome editing efficiency be determined following EnGen® Sau Cas9 (NEB #M0654) transfections or microinjections?
- What is the sequence for a typical guide RNA specific for EnGen® Sau Cas9?
- Which nuclear localization signal is fused to EnGen® Sau Cas9?
- Why do I observe incomplete digestion with EnGen® Sau Cas9 and my target-specific Sau sgRNA in vitro?
- What is the difference between EnGen® Spy Cas9 Nickase (NEB #M0650) and EnGen Spy Cas9 NLS (NEB #M0646)?
- Which nuclear localization signal is fused to EnGen® Spy Cas9 Nickase?
- Does NEB offer synthetic sgRNAs or plasmids for cloning?
- How should offset sgRNAs be oriented for efficient modification using EnGen® Spy Cas9 Nickase?
- Why does binding efficiency differ between two different guide RNAs?
- Does NEB provide plasmids for sgRNA cloning?
- What is the difference between EnGen® Spy Cas9 NLS (NEB #M0646) and Cas9 NLS, S.pyogenes (NEB #M0641)?
- Where is the nuclear localization signal on EnGen® Spy Cas9 NLS located?
- Which nuclear localization signal is fused to EnGen® Spy Cas9 NLS?
- Why do I observe incomplete digestion?
- Does NEB provide plasmids for gRNA cloning?
- Where is the nuclear localization signal on SNAP dCas9 NLS, S. pyogenes located?
- Which nuclear localization signal is fused to SNAP dCas9 NLS, S. pyogenes?
- What inactivating mutations were made in SNAP dCas9 NLS, S. pyogenes?
- Why do I observe incomplete binding with SNAP dCas9 NLS, S. pyogenes?
Protocols for
Programmable Nucleases
- In vitro digestion of DNA with Cas9 Nuclease, S. pyogenes (M0386)
- Protocol for Electroporation of EnGen® SpyCas9 NLS, S. pyogenes RNP (ribonucleoprotein) into adherent cells using the Lonza 4D-Nucleofector™ System
- Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX
- Electroporation of EnGen® Spy Cas9 HF1 (NEB #M0667) into adherent cells using the Lonza 4D-Nucleofector™ System
- Transfection of EnGen® Spy Cas9 HF1 (NEB #M0667) into adherent cells using the Lipofectamine® RNAiMAX System
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CRISPR/Cas9 & Targeted Genome Editing: New Era in Molecular Biology
Understand the history, importance and future of CRISPR/Cas9 and target genome editing
- Genome Editing Brochure
- RNA Synthesis Brochure
Tools & Resources
Feature Articles
Brochures
- Roberts, R.J., Vincze, T., Posfai, J., Macelis, D. (2015) REBASE - A database for DNA restriction and modification: enzymes, genes and genomes Nucleic Acids Res; 43, D298-D299. PubMedID: 25378308
- Morgan, R.D., Luyten, Y.A., Johnson, S.A., Clough, E.M., Clark, T.A. and Roberts, R.J. (2016) Novel m4C modification in type I restriction-modification systems. Nucleic Acids Res; Nov, 2;44(19):9413-9425. PubMedID: 27580720
- Roberts, R.J., Vincze, T., Posfai, J., Macelis, D. (2014) REBASE - A database for DNA restriction and modification: enzymes, genes and genomes Nucleic Acids Res;
Publications related to
Programmable Nucleases
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