Source: Lysinibacillus manganicus An22
500 – 1000 u/ml Store at -20°C
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.
Incubate at 37°C.
1XSEBuffer 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 ZraI, 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 bandsas 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.
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.