Technical Data
M9580
MvaI (EcoRII*)
2000U
Molecular Biology Storage: -20CShipping: Blue Ice
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

Concentration: 10u/ul

Unit Definition:
One unit is defined as the amount of M9580 required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer.

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 37C).

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 37C.

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 4C for short-term only. For long-term storage, aliquot and store at -20C. Stable for at least 12 months
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 37C.

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/1g 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:
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of MvaI for 4 hours.

Thermal Inactivation: Not activated by incubation at 80C for 20min.

Compatible Ends: Satl, Bme1390I

Number of Recognition Sites in DNA:
Lambda: 70
PhiX174: 2
M13mp18/19: 7
pBR322: 6
pUC18/19, pUC57, pTZ19R/U: 5

Protocol for Digestion:
Add:
Nuclease free water: 16ul
10X R1625-04: 2ul
DNA (0.5-1ug/ml): 1ul
M9580: 0.5-2ul
Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours.*

Protocol for Digestion Directly after Amplification:
Add:
PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)
Nuclease free water: 18ul
10X R1625-04: 2ul
M9580: 1-2ul
Mix gently and spin down for a few seconds.
Incubate at 37C for 1-16 hours.*
*Star activity appears at a greater than 5-fold digestion (5ux1hour).
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.