| Recognition site | CCANNNN↑NTGG
GGTN↓NNNNACC |
| Source | Acinetobacter calcoaceticus B7 |
| Assayed on | Lambda DNA (dcm-) |
| Unit definition | One 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-buffer | G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 10 - 25 | 100 | 25 - 50 | 50 - 75 | 50 - 75 | 100 |
|
| Optimal temperature | 37oC |
| 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. |
| Ligation | After 5-fold overdigestion with enzyme 95% 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 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 sensitivity | Blocked by overlapping Dcm methylation (CmCWGG): CCANNNCCTGG or CCAGGNNNTGG. |
| Inactivation 20 minutes under | 65oC |
| Notes | High 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).
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