Sfr274 I

Description

Recognition site and hydrolysis position:

Source: Streptomyces fradiae 274
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (Hind III-digest) in 1 hour at 50°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (Hind III-digest)
Optimal SE-buffer: SE-buffer B
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.. Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 50°C. Reagents Supplied with Enzyme 10 X SE-buffer B.
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by CTCG^mAG methylation.
Not blocked by CT^mCGAG methylation.
Notes: Incubation at 37°C results in 70% activity
References:
Puchkova, L.I., Krivopalova, G.N., Andreeva, I.S., Selina, A.V., Serov, G.D., Rechkunova, N.I., Degtyarev, S.Kh. Izv. Sib. Otd. Akad. Nauk SSSR 1: 32-34 (1990).