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Restriction Endonucleases >
Nicking Endonucleases >
Nt.CviPII |
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Prices are in US dollars and valid only for US orders.
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Recognition Site:
isoschizomers | single letter code
Description:
Nt.CviPII is a nicking endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate. The final product on pUC19 is an array of bands from 25 to 200 bp. CCT is cut less efficiently than CCG and CCA. Some of the CCT sites are not cleaved.
Source:
An E. coli strain that expresses a fusion of Mxe GyrA intein, chitin-binding domain and a truncated form of the Nt.CviPII nicking endonuclease gene from Chlorella virus NYs-1.
Reagents Supplied:
NEBuffer 4 (10X)
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 25% | | NEBuffer 2: | | 100% | | NEBuffer 3: | | 50% | | NEBuffer 4: | | 100% |
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Heat Inactivation:
65°C for 20 minutes
Survival in a Reaction:
Minimum units to digest 1 µg of substrate DNA in 16 hours: 1.00 unit(s)
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 4
Incubate at
37°C.
1X NEBuffer 4:
20 mM Tris-acetate
50 mM potassium acetate
10 mM Magnesium Acetate
1 mM Dithiothreitol
pH 7.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to digest 1 µg of pUC19 DNA resulting in a stable pattern of fragments between 25 and 200 bp in 1 hour at 37°C in a total reaction volume of 50 µl.
Concentration:
3,000 units/ml
Unit Assay Substrate:
pUC19 DNA
Storage Conditions:
20 mM Tris-HCl
100 mM NaCl
50% Glycerol
pH 8.0 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
Usage notes:- Tests have suggested that an exonuclease activity is an inherent part of the enzyme. Thus a one hour incubation time is thought to be the optimal time for most procedures. To run on an electrophoresis gel, add loading dye to a final concentration of 0.4% SDS.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA
and 1 unit of Nt.CviPII incubated for 16 hours at 37ºC
resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
Reagents Sold Separately
NEBuffer 4
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