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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » MvaI (EcoRII*)

MvaI (EcoRII*)

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Specifications

5'-C C^A G G-3'
3'-G G T^C C-5'
Catalog #M9580
NOTEMvaI-neoschizomer of EcoRII, produces DNA fragments that have a
1-base 5'-extension. Unlike EcoRII, MvaI is not blocked by Dcm methylation.
SourceE.coli that carries the cloned mvaIR gene from Micrococcus varians RFL19
Concentration 10u/ul
Unit DefinitionOne 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.
Supplied WithR1625: 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 StabilityMay 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 AssayNo 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 IncubationA 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 AssayAfter 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 InactivationNot activated by incubation at 80°C for 20min.
Compatible EndsSatl, Bme1390I
Number of Recognition Sites in DNA
Lambda70
PhiX1742
M13mp18/197
pBR3226
pUC18/19, pUC57, pTZ19R/U5
Protocol for Digestion
Add
Nuclease free water16ul
10X R1625-042ul
DNA (0.5-1ug/ml)1ul
M95800.5-2ul
Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.*
Protocol for Digestion Directly after Amplification
Add
PCR Reaction Mixture10ul (~0.1-0.5ug of DNA)
Nuclease free water18ul
10X R1625-042ul
M95801-2ul
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).


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