BstNI

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
R0168S     -20       BstNI R0168SVIAL -20 1 x 0.3 ml 10,000 units/ml   NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
R0168L     -20       BstNI R0168LVIAL -20 2 x 0.75 ml 10,000 units/ml   NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X

1. BstNI is an isoschizomer of EcoRII but cuts DNA at a different location. (EcoRII cuts before the two C residues).
2. BstNI produces DNA fragments that have a single-base 5´ extension which are more difficult to ligate than blunt-ended fragments. More efficient ligation can be achieved by using the Quick Ligation Kit (NEB #M2200).
3. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
4. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.
5. Not sensitive to CpG, dcm, or dam methylation.

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
• Dam and Dcm Methylases of E. coli
• 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
• Compatible Cohesive Ends and Generation of New Restriction Sites
• Dam-Dcm and CpG Methylation
• Enzymes with Multiple Recognition Sequences
• Frequencies of Restriction Sites
• Interrupted Palindromes
• 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. Do degenerate recognition sites need to be palindromic?
2. How can this enzyme be inactivated?
3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
4. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
5. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

• 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]

• R0168S_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]

• R0168S_L_v1_0151302
• R0168S_L_v1_0151308
• R0168S_L_v1_0151402
• R0168S_L_v1_0151409
• R0168S_L_v1_0151411
• R0168S_L_v1_0171505
• R0168S_L_v1_0191509
• R0168S_L_v1_0201511
• R0168S_L_v1_0201604
• R0168S_L_v1_0201701
• R0168S_L_v1_0201707
• R0168S_L_v1_0201802
• R0168L_v1_10017681
• R0168S_v1_10011635
• R0168S_v1_10037391
• R0168L_v1_10037389
• R0168S_v1_10052724
• R0168L_v1_10060406
• R0168L_v1_10068452
• R0168S_v1_10070421
• R0168L_v1_10081575
• R0168L_v1_10090038
• R0168S_v1_10095483
• R0168L_v1_10104510
• R0168S_v1_10104241
• R0168L_v1_10118174
• R0168S_v1_10123550
• R0168S_v1_10131170
• R0168L_v1_10148767
• R0168S_v1_10152802
• R0168L_v1_10168858
• R0168S_v1_10168860
• R0168L_v1_10180951
• R0168S_v1_10180953
• R0168L_v1_10215558
• R0168S_v1_10229438
• R0168L_v2_10258640
• R0168S_v2_10258648
• R0168S_v2_10298105
• R0168L_v2_10298107
• R0168S_v2_10324883
• R0168L_v2_10324880

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

• BstNI
• 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.