Description
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
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.
- This product is related to the following categories:
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Restriction Endonucleases: A,
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High-Fidelity (HF) Restriction Endonucleases,
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Time-Saver Qualified Restriction Enzymes,
- This product can be used in the following applications:
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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 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
NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%
Diluent Compatibility
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