Product info: AccB7 I


Name
AccB7 IBlocked by CCANNNN(5mc)TGG or 
C(5mC)ANNNNNTGG methylation   
Cat. #E179E180E179TE180T
Package, u.a.20010002001000
Concentration, u.a./ml5000500050005000

Recognition site
CCANNNNNTGG
GGTNNNNNACC
SourceAcinetobacter calcoaceticus B7
Assayed onLambda DNA (dcm-)
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Optimal SE-bufferG (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Enzyme activity (%)
BGOWYRose
10 - 2510025 - 5050 - 7550 - 75100
Optimal temperature
37oC
Storage conditions10 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.
LigationAfter 5-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme 10 X SE-buffer G
For E179T and E180T an additional 10X SE-buffer ROSE is supplied.
Methylation sensitivityBlocked by overlapping Dcm methylation (CmCWGG): CCANNNCCTGG or CCAGGNNNTGG.
Inactivation 20 minutes under
65oC
NotesHigh enzyme concentration may result in star activity.
Turbo AccB7 I can be used for short time (10 min) DNA digestion as well as for standard reaction. The reaction can be performed using optimal or universal (ROSE) Buffer.
Turbo DNA Digestion:
Applications:
-Fast DNA analysis
-Fast preparation of vectors for cloning
-Double digestion
Enzyme Properties: 1 μl of Turbo AccB7 I cuts 1 μg of DNA in 1 x SE-Buffer G or universal 1 x SE-Buffer ROSE in 10 min (see the protocol below). A short time of DNA digestion requires high quality purification of DNA sample. The enzyme may be used in standard conditions of DNA cleavage (E179).
Turbo reaction protocol: 20 μl of the reaction volume:
10 x SE Buffer ROSE - 2 μl
DNA - 0,2-1 μg
Nuclease-free water - to 20 μl
+ 1μl of Turbo AccB7 I
Incubate at 37°C for 10 min.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Dedkov, V.S., Repin, V.E., Rechkunova, N.I., Degtyarev, S.Kh., Verhosina, V.A.,Vinogradova, T.P, Izv. Sib. Otd. Akad. Nauk SSSR 1: 35-37 (1990).