HindIII

Product Overview

• Article Name: HindIII
• Biozol Catalog Number: USB-H4000
• Supplier Catalog Number: H4000
• Alternative Catalog Number: USB-H4000-5X5000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-A A G C T T-3 3-T T C G A A-5 Source: Haemophilus influenzae Rd Concentration: 10u/ul Unit Definition: One unit is defined as the amount of HindIII required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer 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). R1625-04: Restriction Enzyme Buffer E, 10X: Supplied as a liquid in 10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubate at 37C. Dilution Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 250mM KCl, 0.1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Thermal Inactivation: HindIII is inactivated by incubation at 80C for 20min. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with HindIII. Ligation/Recleavage (L/R) Assay: The ligation and recleavage assay was replaced with LO test after validating experiments showed LO 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 single- stranded or double-stranded labeled oligonucleotide was observed after incubation with 10 units of HindIII for 4 hours. Blue/White Cloning Assay: The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test. Stability during Prolonged Incubation: A minimum of 0.1 units of HindIII is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Number of Recognition Sites in DNA: Lambda: 6: M13mp18/19, pBR322, pUC18/19, pUC57, pTZ19R/U: 1: PhiX174: 0 Digestion of Agarose-embedded DNA: A minimum of 5 units of HindIII is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Methylation Effects: Dam, Dcm, EcoKl, CpG: : never overlaps - no effect EcoBl: may overlap - cleavage impaired. Recommended Protocol for Digestion: Add: Nuclease free water: 16ul R1625-04: 2ul DNA (0.5-1ug/ul): 1ul HindIII: 0.5-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Protocol for Digestion of PCR products directly after amplification: Add: PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA) Nuclease free water: 18ul R1625-04: 2ul HindIII: 1-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Storage and Stability: May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.