E0374-50
Eco91I (BstEII)
Size
£294.00
- SKU:
- E0374-50
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 5'-G↓G T N A C C-3' 3'-C C A N T G↑G-5' Source: Escherichia coli RFL91 Concentration: 10u/ul Supplied With: R1625 - Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate (pH 7.9 at 37°C), 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. R1625-03: Restriction Enzyme Buffer D, 10X: Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA when diluted to 1X. Incubate at 37°C. Unit Definition: One unit is defined as the amount of Eco91I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of recommended reaction buffer. Diluent Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Storage Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Enzyme Properties: Methylation Effects: Dam: never overlaps - no effect. Dcm/CpG: may overlap - no effect. EcoKl/EcoBl: may overlap - effect not determined. Stability during Prolonged Incubation: A minimum of 0.1 units of E0374-50 is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. Thermal Inactivation: E0374-50 is inactivated by incubation at 65°C for 20 minutes. Digestion of Agarose-embedded DNA: A minimum of 5 units of E0374-50 is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Compatible Ends: G↓GT(C/G)ACC–MaeIII, NmuCI G↓GT(A/T)ACC–MaeIII Number of Recognition Sites in DNA: Lambda: 13 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0 Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with Eco911 (10u/ug lambda DNA x 16 hours). Ligation/Recleavage (L/R) Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase 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 Eco911for 4 hours. Blue/White Cloning Assay: The B/W assay was replaced by the L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that equals to that of B/W test. Storage and Stability: Aliquot and store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Recommended Protocol for Digestion: Add: Nuclease-free water 16ul R1625-03 2ul DNA (0.5-1ug/ul) 1ul Eco91I 0.5-2ul* Mix gently and spin down for a few seconds. Incubate at 37°C for 1-16 hours*. The digestion reaction may be scaled either up or down.
- Shipping Conditions:
- Blue Ice