H3999-80
HincII (HindII)
Size
£294.00
- SKU:
- H3999-80
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 5'-G T Py^Pu A C-3' 3'-C A Pu^Py T 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), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with HincII. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with HincII, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.3uM. More than 95% 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. Methylation Effects: GTm5CGAC and the hemi-methylated sequence GTPyPuAm5C are cleaved by HincII at a significantly slower rate. Cleavage is blocked when both strands are methylated as shown: 5'… GTPyPuAm5C…3' 3'…m5CAPuPyT G…5'. Overlapping CG methylation may influence DNA cleavage. HincII does not cut GTPyPum6AC. Blocked by overlapping EcoBI or EcoKI methylation. 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. Number of Recognition Sites in DNA: Lambda: 35 PhiX174: 13 M13mp18/19: 1 pBR322: 2 pUC18/19: 1 pUC57: 1 pTZ19R/U: 1 pBluescriptIIKS(-/+): 1 pBluescriptIISK(-/+): 1 pACYC177: 1 pACYC184: 2 Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
- Shipping Conditions:
- Blue Ice