|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-C C^A G G-3'
3'-G G T^C C-5'
MvaI-neoschizomer of EcoRII, produces DNA fragments that have a
1-base 5'-extension. Unlike EcoRII, MvaI is not blocked by Dcm methylation.
E.coli that carries the cloned mvaIR gene from Micrococcus varians RFL19
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.
Supplied in 10mM Tris-HCl, pH 7.5, 400mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 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 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. For long-term storage, aliquot and store at -20°C. 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 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:
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 80°C for 20min.
Compatible Ends: Satl, Bme1390I
Number of Recognition Sites in DNA:
pUC18/19, pUC57, pTZ19R/U: 5
Protocol for Digestion:
Nuclease free water: 16ul
10X R1625-04: 2ul
DNA (0.5-1ug/ml): 1ul
Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.*
Protocol for Digestion Directly after Amplification:
PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)
Nuclease free water: 18ul
10X R1625-04: 2ul
Mix gently and spin down for a few seconds.
Incubate at 37ºC for 1-16 hours.*
*Star activity appears at a greater than 5-fold digestion (5ux1hour).