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Bal I
Recognition & cleavage sequence
Source: Brevibacterium
albidum
Reaction
conditions
1X
EzBuffer Bal I, 37℃
1X
EzBuffer Bal I
50
mM Tris-HCI (pH 8.2 @ 25℃), 5 mM MgCl2
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℃), 50 mM NaCl,
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
Bal
I can be inactivated at 65℃ for 20 min.
Methylation
sensitivity
dam methylation:
Not sensitive
dcm methylation: Conditionally
sensitive
CpG methylation:
Not sensitive
Relative
activity in EzBuffers
EzBuffer
I: 0%
EzBuffer
II: 75%
EzBuffer
III: 25%
EzBuffer
IV: 75%
FastCut
Buffer: Not recommended
Note
Activity
is inhibited by dcm methylation partially overlapping its recognition
sequence.
● 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 20-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.