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Specifications

5'-A^C G C G T-3'
3'-T G C G C^A-5'
Catalog #	M4140
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
Storage and Stability	May be stored at 4°C for short-term only. For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 6 months 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.
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.
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.