5'-G A C N N NˇN N G T C-3' 3'-C T G N N^N N N C A G-5'
Concentration: 10u/ul
Source: Enterobacter amnigenus RFL1105
Supplied with:R1625-50: Restriction Enzyme Buffer, 10X for E0200-05Supplied in a 10X concentrated stock. Dilute to 1X or 2X before use
When used as a 1X liquid the form is 10mM Tris-HCl, 5mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA. Incubate at 37°C
R1625: Restriction Enzyme Buffer A, 10XSupplied in a 10X concentrated stock. Dilute to 1X or 2X before use
When used as a 1X liquid the form is 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA
Storage Buffer:Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay:No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours).
Ligation/Recutting Assay:The ligation and recutting assay was replaced with LO test ater validating experiments showed LO test ability to trace nuclease and phosphate activities with sensitivity that is higher than L/R by a factor of 100.
Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of E0200-05 for 4 hours.
Star Activity:An excess of enzyme (7.5u/ug DNA x 16 hours) may result in star activity.
Stability during Prolonged Incubation: A minimum of 0.1 units 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 5 units of Eam11051 is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA:Lambda: 9 PhiX174, pTZ19R/U, pBR322, pUC18/19, pUC57: 1M13mp18/19: 0
Enzyme Properties:Methylation Effects on Digestion:Dam, Dcm, EcoKl, EcoBl: never overlaps - no effectCpG: May overlap - no effect
