1000 u/ml
Recognition Sequence:
5’… GCTCCN …3’
3’… CGAGGN …5’
Sourse: Lysinibacillus manganicus An22
Supplied in:
10 mM Tris-HCl (pH 7.4); 50 mM NaCl;
0,1 mM EDTA; 200 µg/ml BSA;
1 mM DTT; 50% glycerol.
Reaction Conditions:
1µSEBuffer B.
Incubate at 37°C.
1´SEBuffer B
10 mM Tris-HCl (pH 7.6 @ 25ºC) 10 mM MgCl2
1 mM DTT
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.
Quality Control Assays
Ligation: 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.
16-Hour Incubation:
A 50 µl reaction containing 1 µg of λ 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.
Oligonucleotide Assay:
No detectable degradation of a single-
and double-stranded oligonucleotide was
observed after incubation with 10 units of
enzyme for 3 hours.
Enzyme Properties
Activity in SEBuffers:
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.
Heat Inactivation:
Yes (80°C for 20 minutes)
Reagents Supplied with Enzyme:
10XSEBuffer B
Notes: High enzyme concentration may result in star activity. The minimum number of units that resulted in complete digestion of 1 µg 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.