Enzima restrictiva de clasa I EcoRI-HF
Caracteristici standard:
| Product Name | EcoRI-HF® |
| Concentration: | 100,000 units/ml |
| Unit Definition: | One unit is defined as the amount of enzyme required to digest 1 µg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 µl. |
| Shelf Life: | 24 months |
| Storage Temp: | -20°C |
| Storage Conditions: | 300 mM NaCl, 10 mM KPOΓ , 1 mM DTT, 0.1 mM EDTA, 50 % Glycerol, 0.15 % TritonX-100, 200 µg/ml BSA, (pH 7.0 @ 25°C) |
| Specification Version: | PS-R3101T/M v2.0 |
| Effective Date: | 16-May-16 |
| Blue-White Screening (Terminal Integrity) | - A sample of pUC19 vector linearized with a 10-fold excess of EcoRI-HF™, religated and transformed into an E. coli strain expressing the LacZ beta fragment gene results in |
| Endonuclease Activity (Nicking) | - A 50 µl reaction in CutSmart™ Buffer containing 1 µg of supercoiled PhiX174 DNA and a minimum of 100 Units of EcoRI-HF™ incubated for 4 hours at 37ºC results in |
| Exonuclease Activity (Radioactivity Release) | - A 50 µl reaction in CutSmart™ Buffer containing 1 µg of a mixture of single and double -stranded [ ³H] E. coli DNA and a minimum of 200 units of EcoRI-HF™ incubated for 4 hours at 37ºC releases |
| Ligation and Recutting (Terminal Integrity) | - After a 20-fold over-digestion of Lambda DNA with EcoRI-HF™, >95% of the DNA fragments can be ligated with T4 DNA ligase in 16 hours at 16ºC. Of these ligated fragments, >95% can be recut with EcoRI-HF™. |
| Non-Specific DNase Activity (16 Hour) | - A 50 µl reaction in CutSmart™ Buffer containing 1 µg of Lambda DNA and a minimum of 100 Units of EcoRI-HF™ incubated for 16 hours at 37ºC results in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis |
| Protein Purity Assay (SDS-PAGE) | - EcoRI-HF™ is >95% pure as determined by SDS PAGE analysis using Coomassie Blue detection |
