Technical Data
M9580
MvaI (EcoRII*)
2000u
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Technical Data |
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M9580 |
MvaI (EcoRII*) |
2000u |
| Molecular Biology | Storage: -20°CShipping: Blue Ice |
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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 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 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. 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: 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 37ºC 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 37ºC for 1-16 hours.* *Star activity appears at a greater than 5-fold digestion (5ux1hour). |
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