|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-C A G^C T G-3'
3'-G T C^G A C-5'
One unit is defined as the amount of PvuI required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Supplied as a liquid in10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
R1625 Restriction Enzyme Buffer A, 10X
Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9
R1625-02 Restriction Enzyme Buffer C, 10X Supplied as a liquid in 10mM Tris-HCl pH 7.5 10mM Magnesium chloride, 50mM sodium chloride, 0.1mg/ml BSA Inhibition Temperature: 37°C.
Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, aliquot and store at -20°C. Stable for at least 12 months
Restriction Enzyme Buffer A is provided to simplify buffer selection for double digests. 98% of Restriction Endonucleased work well in a 1x or 2x concentration of theRestriction Enzyme Buffer A.
Number of Recognition Sites:
M13mp18/19: 3 pBR322: 1
pUC18/19: 2 pUC57 : 2
PvuI is inactivated by incubation at 80°C for 20min.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 10-fold overdigestion (15u/ug lambda DNA x 1 hours) with PvuII (see Star Activity).
Ligation/Recutting Assay: After 10-fold overdigestion (5u/ug DNA x 2 hours) with PvuII, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.06uM. 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 PvuI for 4 hours.
Star Activity: A large excess of enzyme (15u/ug DNA x 1 hours) may result in star activity. Star activity appears at 10-fold overdigestion (10 units x 1 hour).
Methylation Effect: Dam:, Dcm, CpG, EcoKI: never overlaps-no effect
EcoBI: may overlaps- not determined
Stability during Prolonged Incubation: A minimum of 0.2units of PvuI are required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.