NlaIII

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
R0125S     -80       NlaIII R0125SVIAL -80 1 x 0.05 ml 10,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X   Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
R0125L     -80       NlaIII R0125LVIAL -80 1 x 0.25 ml 10,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X   Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X

Materials Sold Separately

• rCutSmart™ Buffer

1. NaCI and KCI inhibit activity, (NH₄)₂ SO₄ does not.
2. Cleaves to leave a 3´ CATG extension which can be ligated to DNA fragments generated by SphI.
3. NlaIII retains full activity for 3 months at -20°C.
4. For long term storage, aliquot and store at -80°C. 
5. The enzyme retains full activity after at least 10 free-thaw cycles; the enzyme can be aliquoted and stored at –80C.
6. Not sensitive to CpG, dcm, or dam methylation.

1. Optimizing Restriction Endonuclease Reactions
2. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
3. Restriction Digest Protocol
4. 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
• Restriction Enzymes for Droplet Digital PCR (ddPCR)
• Single Letter Codes
• Star Activity
• Traditional Cloning Quick Guide

• Alphabetized List of Recognition Sequences
• Compatible Cohesive Ends and Generation of New Restriction Sites
• Dam-Dcm and CpG Methylation
• Frequencies of Restriction Sites
• Isoelectric Points (pI) for Restriction Enzymes
• Isoschizomers
• NEB Diluent and Buffer Table
• Time-Saver™ Qualified Enzymes
• Why Choose Recombinant Enzymes?

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

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

1. Is NlaIII inhibited by salt?
2. What are the storage conditions for NlaIII?
3. Does NlaIII produce commonly used compatible ends?
4. Is extended digestion of NlaIII recommended?
5. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
6. 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?
7. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
8. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
9. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
10. 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]

• R0125S_L_v3

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]

• R0125S_L_v1_0631304
• R0125S_L_v1_0631309
• R0125S_L_v1_0631312
• R0125S_L_v1_0631403
• R0125S_L_v1_0631407
• R0125S_L_v1_0631502
• R0125S_L_v2_0631507
• R0125S_L_v1_0631411
• R0125S_L_v1_0631505
• R0125S_L_v2_0661603
• R0125S_L_v2_0661609
• R0125S_L_v2_0661704
• R0125S_L_v2_0661709
• R0125S_L_v3_0661804
• R0125L_v3_10021477
• R0125S_v3_10021445
• R0125L_v3_10038680
• R0125S_v3_10038682
• R0125L_v3_10055795
• R0125S_v3_10055794
• R0125L_v3_10069830
• R0125S_v3_10077698
• R0125L_v3_10084081
• R0125L_v3_10090026
• R0125S_v3_10090027
• R0125S_v3_10092195
• R0125L_v3_10097402
• R0125L_v3_10114388
• R0125S_v3_10114389
• R0125L_v3_10133537
• R0125S_v3_10140759
• R0125L_v3_10144734
• R0125L_v3_10155077
• R0125S_v3_10155080
• R0125L_v3_10174519
• R0125S_v3_10174520
• R0125L_v3_10181587
• R0125L_v3_10197926
• R0125L_v3_10207386
• R0125S_v3_10207388
• R0125S_v3_10242557
• R0125L_v3_10242577
• R0125L_v3_10260184
• R0125S_v3_10260281
• R0125L_v3_10272769
• R0125L_v3_10288692
• R0125S_v3_10294300
• R0125L_v3_10314034
• R0125S_v3_10330447
• R0125L_v3_10335265

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

• NlaIII
• rCutSmart™ Buffer
• Gel Loading Dye, Purple (6X)

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.