HinfI

Product Overview

• Article Name: HinfI
• Biozol Catalog Number: USB-H4050
• Supplier Catalog Number: H4050
• Alternative Catalog Number: USB-H4050-2000,USB-H4050-5X2000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-G A N T C-3 3-C T N A G-5 Source: Haemophilus influenzae Rf Concentration: 10u/ul Form: Supplied as a liquid in 10mM Tris-HCl (pH 7.5 at 25C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Restriction Buffer A, 10X: (R1625) Supplied as a liquid in 33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37C Restriction Enzyme Buffer E, 10X: (R1625-04) Supplied as a liquid in 10mM Tris-HCl (pH8.5), 10mM MgCl2, 100mM KCl, 0.1mg/ml BSA. Diluent Buffer: Dilute with 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Thermal Inactivation: HinfI is inactivated by incubation at 65C for 20min. Number of Recognition Sites in DNA Lambda: 148 PhiX174: 21 M13mp18/19: 27 pBR322: 10 pUC18/19: 6 pUC57: 5 pTZ19R/U: 9 Unit Definition: One unit is defined as the amount of HinfI required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of reaction buffer. Recommended Protocol for Digestion: Add: Nuclease free water: 16ul 10X Restriction Buffer: 2ul DNA (0.5-1ug/ul): 1ul HinfI: 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 10X Restriction Buffer: 2ul HinfI: 1-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with HinfI (10u/ug lambda DNA x 16 hours) . Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with HinfI, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 1.3uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide occured during incubation with 10 units of HinfI for 4 hours. Methylation Effects: Dam: never overlaps-no effect Dcm: never overlaps-no effect CpG: may overlap-cleavage impaired EcoKI: never overlaps-no effect EcoBI: may overlap-blocked Stability during Prolonged Incubation: A minimum of 0.1 units of HinfI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C.