MluI

Blue Ice

Molecular Biology Grade

5'-A^C G C G T-3'
3'-T G C G C^A-5'

Source:
Microcccus luteus

Concentration:
~10u/ul

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

Incubation Temperature:
37°C

Diluent Buffer:
10mM Tris-HCl, pH 7.4 at 25°C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol, or Storage Buffer.

Storage Buffer:
10mM Tris-HCl, pH 7.5 at 25°C, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol.

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X
R1625-04: Restriction Enzyme Buffer E, 10X

Enzyme Properties:
Methylation Effects:
Dam: Never overlaps - no effect.
Dcm: Never overlaps - no effect.
CpG: Completely overlaps- blocked.
EcoKl: May overlap - no effect.
EcoBl: May overlap - no effect.

Stability during Prolonged Incubation: A minimum of 0.1units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.

Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20 minutes.

Digestion of Agarose-embedded DNA: A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Compatible Ends: AflIII, AscI, DsaI, PauI

Number of Recognition Sites in DNA:
Lambda: 7
PhiX174: 2
M13mp18/19: 0
pBR322: 0
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0

Quality Control:
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with MluI.

Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with MluI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.07uM. More than 95% of these can be recut.

Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.

Blue/White Cloning Assay: The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.