Product SourceAn E. coli strain that carries the CviAII gene from Chlorella virus PBCV-1 (J.L. Van Etten).
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1 µg of pUC19 DNA in 1 hour at 25°C in a total reaction volume of 50 µl.
1X CutSmart® Buffer
Incubate at 25°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 NEBuffersNEBuffer 1.1: 50%
NEBuffer 2.1: 50%
NEBuffer 3.1: 10%
CutSmart® Buffer: 100%
10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
0.15% Triton® X-100
pH 7.4 @ 25°C
Heat Inactivation65°C for 20 min
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
- Exonuclease Activity (Radioactivity Release):
The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
- Ligation and Recutting (Terminal Integrity):
After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.
- Non-Specific DNase Activity (16 hour):
The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
- CviAII is a neoschizomer of NlaIII.
- Is CviAII activity sensitive to dam, dcm or mammalian CpG methylation?
- How many base pairs should be added at the end of a PCR primer after the CviAII recognition site to guarantee that CviAII will cut properly?
- Does CviAII have any neoschizomers?
- My CviAII enzyme was working fine several months ago, but now it is not working anymore. Is there a reason?
- I have run my DNA digested with CviAII in a gel and I cannot see the digested fragment that I was expecting. Has the enzyme degraded my substrate DNA?
- What is the activity of CviAII at 37°C?
- My restriction enzyme used to be available at a lower concentration. Why does it now come at a higher concentration of 10,000 u/ml?
- What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
- 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?
- How can I access the old Double Digest Finder?
- 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?