E0374-20
Eco47III
Size
£385.00
- SKU:
- E0374-20
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- Sequence: 5'-A G C^G C T-3' 3'-T C G^C G A-5' Source: Escherichia coli RFL47 Concentration: ~10u/ul Unit Definition: One unit is defined as the amount of Eco47III required to digest 1ug of lambda DNA-Eco81I fragments in 1 hr at 37°C in 50ul assay buffer. Storage and Diluent Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Supplied with: R1625: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9 at 37ºC. R1625-03: Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA. Incubate at 37°C Double Digests: R1625 is provided to simplify buffer selection for double digests. 98% of Eco47III, works well in a 1x or 2x. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Eco47III. Ligation/Recutting Assay: The ligation and recutting assay was replaced with L0 assay after validating experiments showed L0 assay 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 Eco47III for 4 hours. Blue/White Cloning Assay: The Blue/White assay was replaced by L0 assay after validating experiments showed L0 assay ability to detect nuclease and phosphatase activities with sensitivity that equals to that of Blue/White assay, Methylation Effects: Dam:, Dcm, EcoKI: never overlaps-no effect CpG: completely overlaps- blocked EcoBI: may overlap- not determined Stability during Prolonged Incubation: A minimum of 0.1 units of Eco47III is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. Thermal Inactivation: Eco47III is inactivated by incubation at 65°C for 20 minutes. Digestion of Agarose-embedded DNA: A minimum of 5 units of Eco47III is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda 2 pBR322 4 phiX174, pUC57, pUC18/19, TZ19R/U 0
- Shipping Conditions:
- Blue Ice