85-1379-71　HpaII 1000U　H7030

特徴

• 5'-C^C G G-3' 3'-G G C^C-5'
• Form (1X):Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM sodium chloride, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
• R1625: Restriction Enzyme Buffer A, 10X (for 100% digestion):Supplied as a liquid containing 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
• 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.
• Diluent Buffer:Dilute with 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
• Overdigestion Assay:No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with HpaII (10u/ug lambda DNA x 16 hours) .
• Ligation/Recleavage Assay:The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activites with sensitivity that is higher than L/R by a factor of 100.
• Labeled Oligonucleotide (LO) Assay: No detectable degradation of single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of HpaII for 4 hours.
• Methylation Effects:Dam, Dcm, EcoKI, EcoBI: never overlaps, no effect
• CpG: completely overlaps, blocked
• 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.
• Compatible Ends:Bsp119I, Bsu15I, Hin1I, MaeII, MspI, NarI, Psp1406I, Ssil, TaqI, XmiI
• Number of Recognition Sites in DNA:Lambda: 328PhiX174: 5pBR322: 26pUC57: 13pUC18/19: 13pTZ19R/U: 12M13mp18/19: 18
• Protocol for Digestion:Add: 16ul nudlease free dH2O 2ul Restriction Enzyme Buffer A, 10X 1ul DNA (0.5-1ug/ul) 0.5-2ul HpallMix gently and spin down for a few seconds
• Incubate at 37ºC for 1-16 hours.
• Protocol for Digestion of PCR products Directly after Amplification:Add: 10ul (~0.1-0.5ug DNA) 18ul nuclease free dH2O 2ul Restriction Enzyme Buffer A, 10X 1-2ul HpallMix gently and spin down for a few seconds
• Incubate at 37ºC for 1-16 hours.
• Thermal Inactivation:Enzyme is inactivated by incubation at 65°C for 20 minutes.
• Storage and Stability:May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. 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.