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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » HpaII

HpaII

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Specifications

5'-C^C G G-3'
3'-G G C^C-5'
Catalog #H7030
Concentration 10u/ul
Source: Haemophilus parainfluenzae
Restriction Enzyme Buffer A, 10X (for 100% digestion)33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C.
Diluent Buffer 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or Storage Buffer.
Form Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25°C), 100mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 months 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.
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with HpaII(10u/ug lambda DNA x 16 hours) .
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with HpaII, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 3.1uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours.
Methylation EffectsDam, Dcm, EcoKI, EcoBI: never overlaps, no effect.
CpGCompletely overlaps, blocked
Stability during Prolonged IncubationA minimum of 0.1 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Protocol for Digestion
Add44ul nudlease free dH2O
5ul 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 3ºC for 1-2 hours.
Protocol for Digestion of PCR products Directly after Amplification
Add10ul (~1ug DNA)
16ul nuclease free dH2O
2ul Restriction Enzyme Buffer A, 10X
1-2ul Hpall
Mix gently and spin down for a few seconds.
Incubate at 3ºC for 1-16 hours.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Compatible EndsAciI, Bsp119I, Bsu15I, Hin1I, MaeII, NarI, Psp1406I, TaqI, XmiI
Number of Recognition Sites in DNA
Lambda328
PhiX1745
pBR32226
pUC5713
pUC18/1913
pTZ19R/U12
M13mp18/1918
Unit DefinitionOne 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.


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