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Restriction Endonucleases >
Restriction Endonucleases >
HpaII |
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Recognition Site:
isoschizomers | compatible ends | single letter code
Source:
A E. coli strain that carries the HpaII gene from Haemophilus parainfluenzae (ATCC 49669).
Reagents Supplied:
NEBuffer 1
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 100% | | NEBuffer 2: | | 50% | | NEBuffer 3: | | 10% | | NEBuffer 4: | | 50% |
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Methylation Sensitivity:
| dam methylation: Not sensitive | | dcm methylation: Not sensitive | | CpG methylation: Blocked | |
More information about: Methylation Sensitivity |
Heat Inactivation:
65°C for 20 minutes
Survival in a Reaction:
(+ +) Intermediate activity. Suitable for extended digestion, but < 8 hours.
More information about: Extended Digests with Restriction Enzymes
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 1
Incubate at
37°C.
1X NEBuffer 1:
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.0 @ 25°C
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.
Concentration:
10,000 units/ml and 50,000 units/ml
Unit Assay Substrate:
λ DNA
Storage Conditions:
10 mM Tris-HCl
50 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
General notes:- HpaII is an isoschizomer of MspI.
- Inhibited by salt concentrations > 50 mM KCl.
FAQs
- Is HpaII inhibited by salt?
- What is the activity of HpaII at 25°C?
- Is HpaII affected by methylation?
- Are slow sites found in common vectors?
- What is the molecular weight of HpaII?
- Does spermidine increase HpaII activity?
- Does HpaII cleave ssDNA?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Ligation and Re-cutting:
After a 20-fold overdigestion with HpaII, approximately 75%
of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM)
at 16ºC. Of these ligated fragments, > 95% can be recut with HpaII.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA
and 160 units of HpaII incubated for 16 hours at 37ºC
resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 1,000 units of HpaII with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (105 cpm/μg) for 4 hours at 37ºC
released < 0.05% of the total radioactivity.
Reagents Sold Separately
NEBuffer 1
Legal
Patents:
Invitrogen: Licensed Under U.S. Patent No. 5,248,605
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