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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Mnl I

Mnl I

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Specifications

5'...C C T C (N)7 ...3'
3'...G G A G (N)6 ...5'
Catalog #M4150
SourceE. coli with cloned mnlIR gene from Moraxells nonliquefaciens
Concentration10units/ul
Unit DefinitionOne 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 Temperature37°C
FormSupplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Dilution BufferDilute in the same buffer as Form above.
Supplied withR1625: Restriction Enzyme Buffer A, 10X
Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9 at 37°C, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
R1625-02: Restriction Enzyme Buffer C, 10X:
Dilute to 1X. 1X buffer compostion is 10mM Tris-HCl pH 7.5, 10mM MgCl2, 50mM sodium chloride and 0.1mg/ml BSA. Incubate at 37°C
Storage and StabilityAliquot 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.
Enyme Properties
Methylation Effects
DamNever overlaps - no effect
DcmNever overlaps - no effect
CpGMay overlap - no effect
EcoKINever overlaps - no effect
EcoBIMay overlap - Blocked
Stability during Prolonged IncubationA minimum of 0.5units of enzyme are required for complete digestion of 1ug of DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20 minutes.
Number of Recognition Sites in DNA
Lambda262
PhiX17434
pBR32226
pUC5714
pUC18/1913
pTZ19R/U12
M13mp18/1961
Note
Mnl I produces DNA fragments that have a single base 3-extension which are more difficult to ligate than blunt-ended fragments.
Mnl I may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, heat the digested DNA in the presence of SDS prior to electrophoresis.
Quality Control
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Mnl I.
Ligation/Recutting AssayThe ligation and recleavage assay was replaced with LO test after validating experiment showed LO test ability to trace nuclease phosphatase activities with sensitivity that is hihgher than L/R by a factor of 100.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded or double-stranded labeled oligonucleotide was observed after incubation with 10units of Mnl I for 4 hours.
Important NoteThis 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.


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