Technical Data
Molecular Biology Storage: -20CShipping: Blue Ice
5'-C^C G G-3'
3'-G G C^C-5'

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 37C.

Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 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 25C), 100mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 6 months at -20C. 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 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/Recutting Assay: After 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) Assay: No 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 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 37C.

Protocol for Digestion:
Add: 44ul 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 3C for 1-2 hours.

Protocol for Digestion of PCR products Directly after Amplification:
Add: 10ul (~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 3C for 1-16 hours.

Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min.

Compatible Ends: AciI, Bsp119I, Bsu15I, Hin1I, MaeII, NarI, Psp1406I, 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

Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.