sgRNA Synthesis Reagents and Kits
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FAQs for
sgRNA Synthesis Reagents and Kits
- Is it necessary to add a “G” into the oligo following the T7 promoter sequence?
- The yield of my sgRNA is low – what can I do to improve the yield?
- Is it necessary to DNase treat my sgRNA synthesis reaction?
- Why is my sgRNA not exhibiting 100% in vitro Cas9 cleavage activity?
- Is it necessary to phosphatase treat my sgRNAs?
- The sgRNA is not running as a single band on the gel – why is this happening?
- Will the sgRNAs synthesized from this kit (NEB #E3322S) be recognized by homologs of Cas9 from different organisms?
- Do I include the PAM(NGG) in my sgRNA sequence?
- What is the sequence of the control oligo provided with the EnGen sgRNA Synthesis Kit, S. pyogenes?
- Is the Monarch RNA Cleanup Kit (NEB #T2040) compatible with the EnGen sgRNA Synthesis Kit, S. pyogenes?
- Is it necessary to add the provided DTT to the sgRNA synthesis reaction?
- Does NEB provide plasmids for sgRNA cloning?
Protocols for
sgRNA Synthesis Reagents and Kits
- EnGen® sgRNA Synthesis Kit, S. pyogenes Protocol (E3322)
- Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX
- sgRNA Synthesis Using the HiScribe™ Quick T7 High Yield RNA Synthesis Kit (NEB #E2050)
- How to synthesize a sequence-specific sgRNA for experiments using EnGen® Sau Cas9
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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
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Legal Information
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Videos
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Science in 60 CRISPR/Cas9
Get a high level overview of CRISPR/Cas9 and how it is used in genome editing in our Science in 60 segment.
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Gene Editing 101: A practical guide to genome editing
In this webinar you will learn how to increase editing efficiency by directly introducing Cas9 ribonucleoproteins (RNPs) to cells through electroporation or lipofection. Rapid sgRNA synthesis requiring only a single user-supplied ~55mer single-stranded DNA oligonucleotide is described. Methods for assessing genome editing efficiency will be discussed including T7-endonuclease I-based methods, sequencing-based methods, and in vitro Cas9 digestion.
