NheI

Product Overview

• Article Name: NheI
• Biozol Catalog Number: USB-N2425
• Supplier Catalog Number: N2425
• Alternative Catalog Number: USB-N2425-1000,USB-N2425-2500
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-G C T A G C-3 3-C G A T C G-5 Source: Neisseria mucosa heidelbergensis Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-HindIII fragments in 1 hour at 37C in 50ul of assay buffer. Concentration: 10 units/ul Supplied With: R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37C) when diluted to 1X. Diluent Buffer: 10mM Tris-HCl, (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 8.0 at 25C, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Incubation Temp: 37C Compatible Ends: BcuI, Eco130I, XbaI, XmaJI Number of Recognition Sites in DNA Molecules: Lambda: 1 PhiX174: 0 M13mp18/19:0 pBR322: 1 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0 *Note: Supercoiled plasmids may require up to 10-fold more Nhel for complete digestion than linear DNAs (e.g. 10units are required to cleave 1ug of pBR322 DNA). Thermal Inactivation: NheI is inactivated by incubation at 65C for 20min. Stability During Prolonged Incubation: 0.2units of NheI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Enzyme Properties: Methylation Effects on Digestion: Dam, Dcm, EcoKl, EcoBl: never overlaps - no effect CpG: May overlap - cleavage impaired Digestion of Agarose-embedded DNA: A minimum of 5 units of Nhel is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16hrs Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with Nhel. Ligation Recutting Assay: The ligation and recleavage 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 10units of NheI for 4 hours. Storage and Stability: For long-term storage, store at -20C. Aliquots are stable for 6 months after receipt at -20C. 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.