ApoI-HF^®

Product Information

An E. coli strain that carries the cloned and modified ApoI gene from Arthrobacter protophormiae (C. Polisson).

High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.

NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes. Examples of nuclease contamination studies for some of our HF restriction enzymes are shown below.

Restriction Enzyme Competitor Study: Nuclease Contamination

This product is related to the following categories:

Restriction Endonucleases: A Products,

High-Fidelity (HF®) Restriction Endonucleases Products,

Time-Saver Qualified Restriction Enzymes Products

This product can be used in the following applications:

Fast Cloning: Accelerate your cloning workflows with reagents from NEB,

Restriction Enzyme Digestion

• Reagents Supplied

  The following reagents are supplied with this product:

  NEB https://www.neb.com/en/#	Component Name	Component https://www.neb.com/en/#	Stored at (°C)	Amount	Concentration
  R3566S     -20       ApoI-HF® R3566SVIAL -20 1 x 0.05 ml 20,000 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
  R3566L     -20       ApoI-HF® R3566LVIAL -20 1 x 0.25 ml 20,000 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

• Properties & Usage

  Unit Definition

  One unit is defined as the amount of enzyme required to digest 1 μg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 μl.

  Reaction Conditions

  1X rCutSmart™ Buffer
  Incubate at 37°C

  1X rCutSmart™ Buffer
  50 mM Potassium Acetate
  20 mM Tris-acetate
  10 mM Magnesium Acetate
  100 µg/ml Recombinant Albumin
  (pH 7.9 @ 25°C)

  Activity in NEBuffers

  NEBufferâ„¢ r1.1: 10%
  NEBufferâ„¢ r2.1: 100%
  NEBufferâ„¢ r3.1: 10%
  rCutSmartâ„¢ Buffer: 100%
  

  Diluent Compatibility

  • Diluent B

  Storage Buffer

  10 mM Tris-HCl
  200 mM NaCl
  1 mM DTT
  0.1 mM EDTA
  200 µg/ml Recombinant Albumin
  50% Glycerol
  pH 7.4 @ 25°C

  Heat Inactivation

  80°C for 20 minutes

  Methylation Sensitivity

  dam methylation: Not Sensitive
  dcm methylation: Not Sensitive
  CpG Methylation: Not Sensitive
  

  Isoschizomers

  AcsI
  XapI
  

• Related Products

  Companion Products

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

  Materials Sold Separately

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

• Product Notes

  1. Not sensitive to CpG, dcm, or dam methylation.

Protocols, Manuals & Usage

• Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Restriction Digest Protocol

Tools & Resources

• Selection Charts

  • Isoelectric Points (pI) for Restriction Enzymes
  • NEB Diluent and Buffer Table

FAQs & Troubleshooting

• FAQs

  1. Does EcoRI methylase block ApoI-HF?
  2. What does HF® refer to following the name of a restriction enzyme?
  3. Do degenerate recognition sites need to be palindromic?
  4. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  5. 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?
  6. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  7. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
  9. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?

Citations & Technical Literature

• Citations

  Product Citation Tool

Quality, Safety & Legal

• Quality Control Assays

  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.

• Specifications & Change Notifications

  ApoI-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot https://www.neb.com/en/#10177930. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

  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]

  • R3566S_L_v1
  • R3566S_L_v2

• Certificate of Analysis

  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]

  • R3566S_L_v1_0021602
  • R3566S_L_v1_0021609
  • R3566S_L_v1_0021703
  • R3566S_L_v1_0021711
  • R3566S_L_v1_0021804
  • R3566L_v1_10010502
  • R3566S_v1_10018022
  • R3566S_v1_10031780
  • R3566L_v1_10031778
  • R3566S_v1_10050299
  • R3566L_v1_10053571
  • R3566S_v1_10063403
  • R3566L_v1_10070691
  • R3566S_v1_10082137
  • R3566S_v1_10090021
  • R3566L_v1_10087050
  • R3566L_v1_10094062
  • R3566S_v1_10094064
  • R3566L_v1_10116138
  • R3566S_v1_10126513
  • R3566S_v1_10132928
  • R3566S_v1_10152575
  • R3566L_v1_10157941
  • R3566L_v1_10175739
  • R3566L_v2_10178112
  • R3566S_v2_10178113
  • R3566L_v2_10195657
  • R3566S_v2_10195658
  • R3566L_v2_10237547
  • R3566S_v2_10237544
  • R3566S_v2_10260178

• Safety Data Sheets

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

  • ApoI-HF^®
  • rCutSmart™ Buffer
  • Gel Loading Dye, Purple (6X)

• Legal and Disclaimers

  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.