Pse31 I

Enzyme namePse31 I
PrototypeEco31I
Known analogsEco31I
SKUSE-E603
Turbo versionAvailable
High-concentration versionNot available
Recognition site
5'… GGTCTC(N)1▼ …3'
3'… CCAGAG(N)5▲ …5'
SourcePeribacillus species 31 Definition of Activity Unit One unit of activity is defined as the amount of enzyme capable of hydrolyzing 1 µg of T7 DNA in a 50 µL reaction mixture in SE buffer Y at 37°C for 1 hour. Substrate for Activity Determination T7 DNA
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
25-5075-10010-2525-5010075
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onT7 DNA
Storage conditions10 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 200 µg/mL BSA, 7 mM 2-mercaptoethanol, 50% glycerol; Store at -20°C Ligase Compatibility After hydrolysis with a 10-fold excess of enzyme, more than 90% of DNA fragments are ligated by DNA ligase, and approximately 90% of them can be re-cleaved.
LigationAfter digestion with a 10-fold excess of enzyme, more than 90% of DNA fragments can be ligated by DNA ligase, and approximately 90% of these can be recut.
Nonspecific hydrolysisThe pattern of specific hydrolysis remains unchanged when treating 1 µg of DNA with 20 units of enzyme for 16 hours at 37°C. BSA is not recommended for prolonged incubation.
Methylation sensitivityBlocked 90-95% when overlapping with mCG methylation: 5'-GGTCT(5mC)G-3' and/or 5'-GAGAC(5mC)G-3'. Supplied with the Enzyme 10 x SE Buffer Y, BSA (10 mg/mL). For Turbo packaging, 10 X SE Buffer ROSE is included.
Reagents supplied with enzyme10 X SE-buffer Y, BSA
References
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