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HpyCH4 V
Recognition & cleavage sequence
Source: Helicobacter
pylori CH4
Reaction
conditions
1X EzBuffer
IV, 37℃
1X
FastCut Buffer, 37℃
FastCut
Buffer
Enzynomics restriction enzyme can cut substrate DNA in 5-15 min with FastCut Buffer
1X
EzBuffer IV
20
mM Tris-acetate (pH 7.9 @ 25℃), 50 mM
potassium acetate, 10 mM magnesium acetate, 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 7.4 @ 25℃), 100 mM
NaCl, 1 mM dithiothreitol, 0.1 mM EDTA, 200 μg/ml
BSA, 50% Glycerol.
Dilution
buffer: EzDiluent A
10
mM Tris-HCl (pH 7.4 @ 25℃), 50 mM KCl,
0.1 mM EDTA, 1 mM dithiothreitol, 200 ㎍/ml BSA, 50% glycerol.
Heat
Inactivation
HpyCH4
V 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: 25%
EzBuffer
IV: 100%
FastCut
Buffer: 100%
Note
It
is not affected by dam, dcm, or mammalian CpG methylation.
● Material Safety Data Sheet
- Enzynomics_MSDS_R094_HpyCH4 V
● 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 for1 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.