EheI (NarI*)
Référence E2216-500U
Conditionnement : 500U
Marque : US Biological
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E2216 EheI (NarI*)
Grade
Molecular Biology GradeShipping Temp
Blue IceStorage Temp
-20°C
5'-G G C^G C C-3'
3'-C C G^C G G-5'
Concentration:
10u/ul
Buffer:
33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C
Diluent Buffer:
10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or Storage Buffer.
Storage Buffer:
10mM Tris-HCl (pH 7.5 at 25°C), 100mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Applications
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with EheI (see Star Activity). ||Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with EheI, more than 80% of the pBR322 DNA fragments can be ligated in a reaction mixture containing 20-40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.35uM. More than 90% of these can be recut. ||Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours.||Star Activity: A large excess of enzyme (10u/ug DNA x 16 hours) may result in star activity. ||Methylation Effects: EheI does not cut GGm5CGCC. Blocked by CG methylation. ||Stability during Prolonged Incubation: A minimum of 1.0 unit of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. ||Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20min. ||Digestion of Agarose-embedded DNA: A minimum of 20 units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. ||Number of Recognition Sites in DNA:||Lambda: 1|PhiX174: 2|M13mp18/19: 1|pBR322: 4|pUC18/19: 1|pUC57: 1|pTZ19R/U: 0|pBluescriptIIKS(-/+): 0|pBluescriptIISK(-/+): 0|pACYC177: 0|pACYC184: 4||Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-PstI fragments in 1 hour at 37°C in 50ul of assay buffer.