EnGen^® Sau Cas9

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
NEB # Stored at (°C) M0654T -20 Component Name Component # Stored at (°C) Amount Concentration EnGen® Sau Cas9 M0654TVIAL -20 1 x 500 pmol 20 µM NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X

1. 20 µM is equal to ~2.54 mg/ml EnGen^® Sau Cas9.  1 µM is equal to ~0.127 mg/ml.

1. How to synthesize a sequence-specific sgRNA for experiments using EnGen^® Sau Cas9
2. In vitro digestion of DNA with Cas9 Nucleases (NEB #M0386, #M0646, #M0650, #M0654, #M0667, #M0668)

1. How can the genome editing efficiency be determined following EnGen^® Sau Cas9 (NEB #M0654) transfections or microinjections?
2. Which nuclear localization signal is fused to EnGen^® Sau Cas9?
3. Can the EnGen^® sgRNA Synthesis Kit (NEB #E3322S) be used to synthesize sgRNAs specific for Sau Cas9?
4. What is the sequence for a typical guide RNA specific for EnGen^® Sau Cas9?
5. How can I synthesize a sequence-specific sgRNA for experiments using EnGen^® Sau Cas9?
6. Why do I observe incomplete digestion with EnGen^® Sau Cas9 and my target-specific Sau sgRNA in vitro?
7. How do I dilute the enzyme to 1 μM for in vitro reactions?
8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

• Yourik, P., Fuchs, R.T., Mabuchi, M., Curcuru, J.L., Robb, G.B. (2019) Staphylococcus aureus Cas9 is a multiple-turnover enzyme. RNA; [Epub ahead of print], PubMedID: 30348755, DOI: 10.1261/rnr.067355.118

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• M0654S_v1
• M0654T_v1

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• M0654S_v1_10040102
• M0654T_v1_10040099
• M0654T_v1_10052311
• M0654T_v1_10056912
• M0654S_v1_10060288
• M0654T_v1_10065477
• M0654T_v1_10102993
• M0654S_v1_10107281
• M0654S_v1_10119919
• M0654T_v1_10121808
• M0654T_v1_10129877
• M0654S_v1_10132312
• M0654T_v1_10160292
• M0654S_v1_10171037
• M0654T_v1_10182265
• M0654T_v1_10230390
• M0654T_v1_10262789
• M0654T_v1_10305657
• M0654T_v1_10351661

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• EnGen^® Sau Cas9
• NEBuffer™ r3.1

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.