Zsp2 I

Enzyme name Zsp2 I
Prototype NsiI
Known analogs EcoT22I, Mph1103I, NsiI, BfrBI
SKU SE-E145
Turbo version Not available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… ATGCA▼T …3'
3'… T▲ACGTA …5'
Source Zoogloea species 2
Optimal buffer SE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
1005025252550
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA
Storage conditions 10 mM Tis-HCl (pH 7.5); 200 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 20-fold overdigestion with enzyme 90% of the DNA fragments can be ligated and recut
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 37°C
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer B, BSA
Notes To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  1. Unpublished data Dedkov, V.S., Degtyarev, S.Kh unpublished observations.
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