HpaII
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.
Source
Haemophilus parainfluenzae
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
