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Mnl I
Recognition & cleavage sequence
Source: Moraxella
nonliquefaciens
Reaction
conditions
1X
EzBuffer II, 37℃
1X FastCut
Buffer, 37℃
FastCut
Buffer
Enzynomics restriction enzyme can cut substrate DNA in 5-15 min with FastCut Buffer
1X
EzBuffer II
10
mM Tris-HCl (pH 7.9 @ 25℃), 50 mM NaCl,
10 mM MgCl2, 100 μg/ml
BSA
Unit
definition
One
unit is defined as the amount of enzyme required for complete digestion of 1-μg
bacteriophage λ at 37℃ for 1 hr in 50-μl reaction mixtures.
Storage
10
mM Tris-HCl (pH 7.5 @ 25℃), 200 mM KCl,
0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml
BSA, 50% glycerol.
Dilution
buffer: EzDiluent B
10
mM Tris-HCl (pH 7.4 @ 25℃), 300 mM
NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml
BSA, 50% glycerol.
Heat
Inactivation
Mnl
I can be inactivated at 65℃ for 20 min.
Methylation
sensitivity
dam methylation:
Not sensitive
dcm methylation: Not
sensitive
CpG methylation:
Not sensitive
Prolonged
incubation
A
minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 0.25
U.
Relative
activity in EzBuffers
EzBuffer
I: 75%
EzBuffer
II: 100%
EzBuffer
III: 75%
EzBuffer
IV: 100%
FastCut
Buffer: 100%
Note
It
produces a 3’ extension of one nucleotide, which is more difficult to be
ligated than blunt-ends. It is not affected by dam, dcm, or
mammalian CpG methylation.
● Material Safety Data Sheet
● Quality control
Overdigestion assay
DNA digested for 16 hr in 50-μl reaction
with 100 U of enzyme resulted in same the DNA band patterns as those obtained
with 1 U of enzyme for 1 hr.
Ligation and recutting
More than 95% of DNA fragments (1 μg)
digested with 50-fold excess enzyme can be ligated by T4 DNA ligase (400 U) at
16℃ for 16 hr. Of the ligation
products, > 95% can be re-cut.
Extreme pure (EP)
No detectable degradation of 32P-end
labeled single-stranded and double-stranded (5’-, 3’-overhang and blunt end)
oligonucleotides occurred during incubation with 100 U of enzyme for 4 hr.