I-SceI

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
R0694S     -80       I-SceI R0694SVIAL -80 1 x 0.1 ml 5,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X   pGPS2 NotI-linearized Control Plasmid N0420SVIAL -20 1 x 0.01 ml 0.5 mg/ml
R0694L     -80       I-SceI R0694LVIAL -80 1 x 0.5 ml 5,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X   pGPS2 NotI-linearized Control Plasmid N0420SVIAL -20 1 x 0.01 ml 0.5 mg/ml

1. Homing endonucleases do not have stringently-defined recognition sequences in the way that restriction enzymes do. That is, single base changes do not abolish cleavage but reduce its efficiency to variable extents. The precise boundary of required bases is generally not known. The recognition sequence listed is one site that is known to be recognized and cleaved.
2. Supplied with plasmid DNA. NotI-linearized pGPS2 is supplied at 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0), 1 mM EDTA. Cleavage of this 2499 bp plasmid with I-SceI gives fragments of 1518 and 981 base pairs.
3. The enzyme retains full activity after at least 10 free-thaw cycles; the enzyme can be aliquoted and stored at –80C.

1. Optimizing Restriction Endonuclease Reactions
2. Restriction Digest Protocol
3. Double Digest Protocol with Standard Restriction Enzymes

• Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
• Activity of Restriction Enzymes in PCR Buffers
• Cleavage Close to the End of DNA Fragments
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Heat Inactivation
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• Optimizing Restriction Endonuclease Reactions
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Single Letter Codes
• Star Activity
• Traditional Cloning Quick Guide

• Alphabetized List of Recognition Sequences
• Dam-Dcm and CpG Methylation
• Enzymes with Nonpalindromic Sequences
• Frequencies of Restriction Sites
• Isoelectric Points (pI) for Restriction Enzymes
• Isoschizomers
• NEB Diluent and Buffer Table
• Why Choose Recombinant Enzymes?

• Competitor Cross-Reference Tool
• DNA Sequences and Maps Tool

• Double Digest Finder
• Enzyme Finder
• NEBcutter™ v3.0
• NEBioCalculator®
• REBASE®

1. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
2. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
3. For enzymes stored at -80°C, is enzyme activity affected after multiple freeze / thaw cycles?

• Restriction Enzyme Troubleshooting Guide

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]

• R0694S_L_v2

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]

• R0694S_L_v1_0121405
• R0694S_L_v1_0111302
• R0694S_L_v1_0111306
• R0694S_L_v1_0111310
• R0694S_L_v1_0111312
• R0694S_L_v1_0121411
• R0694S_L_v1_0121504
• R0694S_L_v1_0131504
• R0694S_L_v1_0131509
• R0694S_L_v1_0131605
• R0694S_L_v1_0141605
• R0694S_L_v1_0201804
• R0694L_v1_10008634
• R0694L_v1_10022297
• R0694S_v1_10030587
• R0694L_v1_10043034
• R0694S_v1_10050598
• R0694S_v1_10061114
• R0694L_v1_10061113
• R0694S_v1_10072122
• R0694L_v1_10079891
• R0694S_v1_10081460
• R0694L_v1_10084444
• R0694L_v1_10091233
• R0694S_v1_10091197
• R0694S_v1_10092658
• R0694S_v1_10104689
• R0694L_v1_10126310
• R0694S_v1_10126315
• R0694L_v1_10144821
• R0694S_v1_10144823
• R0694L_v2_10156853
• R0694S_v2_10156854
• R0694L_v2_10179261
• R0694S_v2_10179263
• R0694S_v2_10188088
• R0694L_v2_10208330
• R0694S_v2_10208329
• R0694S_L_v1_0201711
• R0694L_v2_10236435
• R0694S_v2_10236434
• R0694L_v2_10253545
• R0694S_v2_10262173
• R0694L_v2_10276590
• R0694S_v2_10299385
• R0694L_v2_10316411
• R0694S_v2_10323131
• R0694L_v2_10338263

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

• I-SceI
• rCutSmart™ Buffer
• pGPS2 NotI-linearized Control Plasmid

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.