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Restriction Endonucleases >
Restriction Endonucleases >
CviKI-1 |
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Prices are in US dollars and valid only for US orders.
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Recognition Site:
isoschizomers | compatible ends | single letter code
Description:
CviKI-1 is a restriction endonuclease with 4 expected recognition sites as well as up to eleven relaxed non-cognate sites (star sites).
Source:
An E. coli strain carrying the cloned and modified CviKI restriction gene derived from CA-1A, a Chlorella virus.
Reagents Supplied:
NEBuffer 4
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 10% | | NEBuffer 2: | | 50% | | NEBuffer 3: | | 25% | | 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:
80°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 pBR322 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.
Concentration:
5,000 units/ml
Unit Assay Substrate:
pBR322 DNA
Storage Conditions:
10 mM Tris-HCl
50 mM NaCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.5 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
General notes:- CviKI-1 overnight digestion or addition of 20% DMSO greatly enhances the star activity. DNA can be digested to small oligos under "star" conditions.
- When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
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 10-fold overdigestion with CviKI-1, > 95%
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 CviKI-1.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA and 10 units of enzyme incubated for 16 hours resulted in the same pattern of DNA bands as a reaction incubated for 1 hour with 1 unit of enzyme.
Reagents Sold Separately
NEBuffer 4
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