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

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 37C in 50ul of assay buffer.

Incubation Temperature: 37C

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

Storage Buffer: 10mM Tris-HCl, pH 7.5 at 25C, 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

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, aliquot and store at -20C. Aliquots are stable for at least 6 months at -20C. 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.
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 37C.

Thermal Inactivation: Enzyme is inactivated by incubation at 65C 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.

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.