SibEnzyme restriction enzymes database

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Acu I

Enzyme name Acu I
Prototype Eco57I
SKU SE-E451
Turbo version Not available
High-concentration version Not available
Recognition site
5'… CTGAAG(N)16▼ …3'
3'… GACTTC(N)14▲ …5'
Source An. E.coli strain that carries the cloned Acu I gene from Acinetobacter calcoaceticus SRW4
Optimal buffer SE-buffer Y + SAM + BSA
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
2550507510050
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 Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligation After 2-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated. Of these, 80% can be recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity not tested
Supplied with enzyme 10 X SE-buffer Y, BSA, SAM
Notes High enzyme concentration may result in star activity. To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml and SAM should be added to a final concentration 10 μM. It is possible to use a final SAM concentration of 10-64 μM. In this case 32 mM SAM solution should be diluted in 3200-500 times, correspondingly. For the small volume of reaction mix (10-20 μl) the stock SAM solution may be previously diluted up to 1 mM (in 32 times) with 5 mM solution of H 2 SO 4 or sterile water and stored at -20°C. Do not use BSA for long incubation.
References
  1. Degtyarev, S.Kh., Kileva, E.V., Dedkov, V.S. Unpublished observations (2001).

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Acu I

Enzyme name Acu I
Prototype Eco57I
SKU SE-E451
Turbo version Not available
High-concentration version Not available
Recognition site
5'… CTGAAG(N)16▼ …3'
3'… GACTTC(N)14▲ …5'
Source An. E.coli strain that carries the cloned Acu I gene from Acinetobacter calcoaceticus SRW4
Optimal buffer SE-buffer Y + SAM + BSA
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
2550507510050
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 Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligation After 2-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated. Of these, 80% can be recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity not tested
Supplied with enzyme 10 X SE-buffer Y, BSA, SAM
Notes High enzyme concentration may result in star activity. To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml and SAM should be added to a final concentration 10 μM. It is possible to use a final SAM concentration of 10-64 μM. In this case 32 mM SAM solution should be diluted in 3200-500 times, correspondingly. For the small volume of reaction mix (10-20 μl) the stock SAM solution may be previously diluted up to 1 mM (in 32 times) with 5 mM solution of H 2 SO 4 or sterile water and stored at -20°C. Do not use BSA for long incubation.
References
  1. Degtyarev, S.Kh., Kileva, E.V., Dedkov, V.S. Unpublished observations (2001).