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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » MlsI (BalI)

MlsI (BalI)

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Specifications

5'-T G G^C C A-3'
3'-A C C^G G T-5'
Catalog #M4130
Source Micrococcus luteus Ng 16-122
Concentration 5u/ul
Unit DefinitionOne unit is defined as the amount of MlsI required to digest 1ug of lambda DNA dcm in 1 hour at 37°C in 50ul of reaction buffer.
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.
Storage and Dilution BufferSupplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Thermal InactivationMlsI is inactivated by incubation at 65°C for 20min.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with MlsI.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with MlsI, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.9uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of single- stranded or double-stranded labeled oligonucleotide was observed after incubation with 10 units of MlsI for 4 hours.
Stability during Prolonged Incubation
A minimum of 0.5units of MlsI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Number of Recognition Sites in DNA
Lambda18
PhiX1740
M13mp18/191
pBR3221
pUC18/190
pUC570
pTZ19R/U0
Digestion of Agarose-embedded DNAA minimum of 20 units of MlsI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Recommended Protocol for Digestion
Add
Nuclease free water16ul
R1625-042ul
DNA (0.5-1ug/ul)1ul
MlsI0.5-2ul
Mix gently, spin down for a few seconds.
Incubate at 37ºC for 1-2 hours.
Protocol for Digestion of PCR products directly after amplification
Add
PCR Reaction Mixture10ul (~0.1-0.5ug of DNA)
Nuclease free water18ul
R1625-042ul
MlsI1-2ul
Mix gently, spin down for a few seconds.
Incubate at 37ºC for 1-16 hours.
Storage and Stability
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.


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