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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
Form10mM Tris-HCl, pH 7.4 at 25°C, 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
R1625-02: Restriction Enzyme Buffer C, 10X
Note: R1625-02 gives 100% digestion where R1625 will only give ~20-50% digestion at the same concentrati
Storage and StabilityAliquot to avoid repeated freezing and thawing. 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.
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 lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
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 AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with Mnl I, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 2uM. More than 90% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease 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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