Companion Products

• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

1. The enzyme retains full activity after at least 10 free-thaw cycles; the enzyme can be aliquoted and stored at –80C.
2. 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.
3. 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.

1. My enzyme used to come with another NEBuffer, but CutSmart™ Buffer is now recommended?  Why?
2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
3. How can I access the old NEBuffer Activity Chart?
4. 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?
5. Why is my Restriction Enzyme not cutting DNA?
6. Why do I see a DNA smear on an agarose gel after a restriction digest?
7. Why do I see additional DNA bands on my gel after a restriction digest?
8. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?

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

• Alphabetized List of Recognition Specificities
• Buffer and Diluent Formulation Table
• Cross Index of Recognition Sequences
• Dam-Dcm and CpG Methylation
• Enzymes with Nonpalindromic Sequences
• Frequencies of Restriction Sites
• Isoschizomers
• Why Choose Recombinant 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
• Megabase Mapping
• 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

• Restriction Enzyme Troubleshooting Guide

• DNA Sequences and Maps Tool

• Double Digest Finder
• Enzyme Finder
• NEBcutter V2.0
• NEBioCalculator
• REBASE

• R0694S_L_v1

• 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

• I-SceI
• CutSmart® Buffer
• pGPS2 NotI-linearized Control Plasmid