Home Restriction Endonucleases Products DraIII-HF®

DraIII-HF®

Dra-III-cutsite_1

Isoschizomers | Single Letter Code

High-Fidelity (HF^®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (CutSmart^® Buffer). All HF-restriction enzymes come with Gel Loading Dye, Purple (6X). Enjoy the enhanced performance and added value of our engineered enzymes at the same price as the native enzyme:

Engineered for improved performance
100% activity in CutSmart Buffer
Time-Saver™ qualified for digestion in 5-15 minutes
Reduced star activity
Supplied with 1 vial of Gel Loading Dye, Purple (6X)

Product Information
High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart 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

EcoRI, NotI, and BamHI from multiple suppliers were tested in reactions containing a fluorescent labeled single stranded, double stranded blunt, 3’overhang or 5’ overhang containing oligonucleotides. The percent degradation is determined by capillary electrophoresis and peak analysis. The resolution is at the single nucleotide level.

Product Source
An E. coli strain that carries the cloned and modified DraIII gene from Deinococcus radiophilus (ATCC 27603).
Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration

Gel Loading Dye, Purple (6X) 25 6X

CutSmart® Buffer -20 10X

Product Categories:

Restriction Endonucleases C G,

High-Fidelity (HF®) Restriction Endonucleases Products,

time-saver-qualified-restriction-enzymes Products

Applications:

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

Restriction Enzyme Digestion
- 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 CutSmart® Buffer
  Incubate at 37°C
  
  1X CutSmart® Buffer
  50 mM Potassium Acetate
  20 mM Tris-acetate
  10 mM Magnesium Acetate
  100 µg/ml BSA
  (pH 7.9 @ 25°C)
  
  Activity in NEBuffers
  CutSmart® Buffer: 100%
  NEBuffer™ 1.1: 10%
  NEBuffer™ 2.1: 50%
  NEBuffer™ 3.1: 10%
  
  Diluent Compatibility
  
  Diluent B
  
  Storage Temperature
  -20°C
  Storage Conditions
  
  10 mM Tris-HCl
  300 mM NaCl
  1 mM DTT
  0.1 mM EDTA
  200 µg/ml BSA
  50% Glycerol
  pH 7.4 @ 25°C
  
  Heat Inactivation
  No
  Methylation Sensitivity
  
  dam methylation: Not Sensitive
  dcm methylation: Not Sensitive
  CpG Methylation: Impaired by Overlapping
- Related Products
  Companion Products
  
  Monarch® Plasmid Miniprep Kit
  
  Monarch® DNA Gel Extraction Kit
  
  Monarch® PCR & DNA Cleanup Kit (5 μg)
  
  Materials Sold Separately
  
  Gel Loading Dye, Purple (6X)
  
  CutSmart® Buffer
- Product Notes
  Cleavage of mammalian genomic DNA is impaired by some combinations of overlapping CpG methylation.
Protocols, Manuals & Usage
- Protocols
  Optimizing Restriction Endonuclease Reactions
  
  Double Digest Protocol with Standard Restriction Enzymes
  
  Restriction Digest Protocol
- Usage Guidelines
  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
  
  Effects of CpG Methylation on Restriction Enzyme Cleavage
  
  Heat Inactivation
  
  Megabase Mapping
  
  NEBuffer Activity/Performance Chart with Restriction Enzymes
  
  Optimizing Restriction Endonuclease Reactions
  
  Reduced Star Activities of HF® Enzymes
  
  Restriction Endonucleases - Survival in a Reaction
  
  Restriction Enzyme Diluent Buffer Compatibility
  
  Restriction Enzyme Tips
  
  Single Letter Codes
  
  Star Activity
  
  Traditional Cloning Quick Guide
Tools & Resources
- Selection Charts
  Alphabetized List of Recognition Specificities
  
  Buffer and Diluent Formulation Table
  
  Compatible Cohesive Ends and Generation of New Restriction Sites
  
  Cross Index of Recognition Sequences
  
  Dam-Dcm and CpG Methylation
  
  Enzymes with Multiple Recognition Sequences
  
  Frequencies of Restriction Sites
  
  Interrupted Palindromes
  
  Isoschizomers
  
  Time-Saver™ Qualified Enzymes
  
  Why Choose Recombinant Enzymes?
- Web Tools
  Competitor Cross-Reference Tool
  
  DNA Sequences and Maps Tool
  
  Double Digest Finder
  
  Enzyme Finder
  
  NEBcutter V2.0
  
  NEBioCalculator
  
  REBASE
FAQs & Troubleshooting
- FAQs
  What does HF® refer to following the name of a restriction enzyme?
  
  Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  
  How can I access the old NEBuffer Activity Chart?
  
  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?
  
  Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR® and GelRed™ during gel electrophoresis?
  
  Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
  
  Why is my Restriction Enzyme not cutting DNA?
  
  Why do I see additional DNA bands on my gel after a restriction digest?
  
  How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  
  Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
- Troubleshooting
  Restriction Enzyme Troubleshooting Guide
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
  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]
  
  R3510S_L_v1
- 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]
  
  R3510S_L_v1_0021303
  
  R3510S_L_v1_0021403
  
  R3510S_L_v1_0021309
  
  R3510S_L_v1_0021502
  
  R3510S_L_v1_0021507
  
  R3510S_L_v1_0031601
  
  R3510S_L_v1_0031606
  
  R3510S_L_v1_0031701
  
  R3510S_L_v1_0031704
  
  R3510S_L_v1_0031711
  
  R3510S_L_v1_0031805
  
  R3510S_v1_10011444
  
  R3510L_v1_10030802
  
  R3510S_v1_10030799
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  DraIII-HF®
  
  Gel Loading Dye, Purple (6X)
  
  CutSmart® Buffer
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
  
  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.

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