H7030
HpaII
Size
£284.00
- SKU:
- H7030
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 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: 328 PhiX174: 5 pBR322: 26 pUC57: 13 pUC18/19: 13 pTZ19R/U: 12 M13mp18/19: 18 Protocol for Digestion: Add: 16ul nudlease free dH2O 2ul Restriction Enzyme Buffer A, 10X 1ul DNA (0.5-1ug/ul) 0.5-2ul Hpall Mix 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 Hpall Mix 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.
- Shipping Conditions:
- Blue Ice