High enzyme concentration and using of nonoptimal buffer may result in star activity. Do not use BSA for long incubation. To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Sources | Recombinant E. coli strain that carries the cloned ecoRIR gene from Escherichia coli. |
Storage conditions | 10 mM potassium phosphate (pH 7.4 at 25°C), 300 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.2 mg/mL BSA, 0.15% Triton X-100 and 50% glycerol. |
Optimal buffer(10X) | 500 mM Tris-HCl (pH 7.5), 100 mM MgCl2, 1000 mM NaCl, 0.2% Triton X-100, 1 mg/mL BSA. |
Ligations | After 10-fold over digestion with enzyme 95% of the DNA fragments can be ligated and recut. |
Methylation Effects: | Not dam methylation-sensitive, not dcm methylation-sensitive. CpG methylation-sensitive |
Campatible Ends | Xap I, Mun I, Tas I, Acs I. |
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