Technical Data
MlsI (BalI)
Molecular Biology Storage: -20CShipping: Blue Ice
5'-T G G^C C A-3'
3'-A C C^G G T-5'

Source: Micrococcus luteus Ng 16-122

Concentration: 5u/ul

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

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.

Storage and Dilution Buffer:
Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.

Thermal Inactivation:
MlsI is inactivated by incubation at 65C 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) Assay: No 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 37C.

Number of Recognition Sites in DNA:
Lambda: 18
PhiX174: 0
M13mp18/19: 1
pBR322: 1
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0

Digestion of Agarose-embedded DNA: A minimum of 20 units of MlsI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Recommended Protocol for Digestion:
Nuclease free water: 16ul
R1625-04: 2ul
DNA (0.5-1ug/ul): 1ul
MlsI: 0.5-2ul
Mix gently, spin down for a few seconds.
Incubate at 37C for 1-2 hours.

Protocol for Digestion of PCR products directly after amplification:
PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)
Nuclease free water: 18ul
R1625-04: 2ul
MlsI: 1-2ul
Mix gently, spin down for a few seconds.
Incubate at 37C for 1-16 hours.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.