Source:E.colistrain that carries the cloned Zra I gene from Zoogloea ramigera
10 mMTris-HCl (pH 7.4); 50 mM NaCl;
0,1 mMEDTA;200µg/ml BSA;
1 mM DTT; 50% glycerol.
10 mMTris-HCl (pH 7.6 @ 25ºC) 10 mM MgCl2
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.
Quality Control Assays
After 10-fold overdigestion with Zra I,approximately 90% of λ DNA fragments can be ligated with T4 DNA
Ligase and recut. In the presence of 10% PEG ligation is better.
A 50µl reaction containing 1 µg of λ
and 10 units of enzyme incubated for 16 hours
resulted in the same pattern of DNA bands
as a reaction incubated for 1 hour.
No detectable degradation of a single-
and double-stranded oligonucleotide was
observed after incubation with 10 units of
enzyme for 3 hours.
SEBuffer B 100%
SEBuffer G 50-75%
SEBuffer O 25-50%
SEBuffer W 25-50%
SEBuffer Y 75-100%
SEBuffer ROSE 100%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more
enzyme to achieve complete digestion.
Zra I is a neoschizomer of Aat II.
Yes(80°C for 20
Reagents Supplied with
Notes:High enzyme concentration may result in star activity. The minimum number of units that resulted in complete digestion of 1
ug of substrate DNA in 16 hours is 0,5. Zra I cleaves linear plasmid DNA at a rate 1,5-2 times higher than supercoiled plasmid DNA.