5'-C C^A G G-3' 3'-G G T^C C-5'
NOTE
MvaI-neoschizomer of EcoRII, produces DNA fragments that have a 1-base 5'-extension. Unlike EcoRII, MvaI is not blocked by Dcm methylation.
Source
E.coli that carries the cloned mvaIR gene from Micrococcus varians RFL19
Unit Definition
One unit is defined as the amount of M9580 required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Form
Supplied in 10mM Tris-HCl, pH 7.5, 400mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Supplied With
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 at 37°C).
R1625-04: Restriction Enzyme Buffer E, 10X: Supplied as a liquid in 10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubate at 37°C.
Dilute with
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
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.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with M9580.
Stability during Prolonged Incubation
A minimum of 0.1 units of MvaI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Ligation/Recutting Assay
After 10-fold overdigestion (0.6u/ug DNA x 17 hours) with M9580, more than 90% of the DNA fragments can be ligated in a reaction mixture containing 20-40u of T4 DNA ligase/1µg of fragments and 10% PEG at a 5'-termini concentration of 0.6uM. More than 90% of these sites can be recut.
Labeled Oligonucleotide (LO) Assay
Thermal Inactivation
Not activated by incubation at 80°C for 20min.
Compatible Ends
Satl, Bme1390I
Number of Recognition Sites in DNA
pUC18/19, pUC57, pTZ19R/U
5
Protocol for Digestion Directly after Amplification
PCR Reaction Mixture
10ul (~0.1-0.5ug of DNA)
TDS
NCBI
UniProt
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