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BsmB I (Esp3 I)
Recognition & cleavage sequence
Source: Bacillus
stearothermophilus B61
Isoschizomer : Esp3 I
Reaction
conditions
1X
EzBuffer III, 55℃
1X
FastCut Buffer, 55℃
FastCut
Buffer
Enzynomics restriction enzyme can cut substrate DNA in 5-15 min with FastCut Buffer
1X
EzBuffer III
50
mM Tris-HCl (pH 7.9 @ 25℃), 100 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 55℃ for 1 hr in 50-μl reaction mixtures.
Storage
10
mM Tris-HCl (pH 7.4 @ 25℃), 300 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
BsmB
I can be inactivated at 80℃ for 20 min.
Methylation
sensitivity
dam methylation:
Not sensitive
dcm methylation: Not
sensitive
CpG methylation:
sensitive
Prolonged
incubation
A
minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 0.5
U.
Relative
activity in EzBuffers
EzBuffer
I: 10%
EzBuffer
II: 50%
EzBuffer
III: 100%
EzBuffer
IV: 25%
FastCut
Buffer: 100%
Note
It
is an isoschizomer of Esp3 I. Cleavage of mammalian genomic DNA is blocked by
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.
Endonuclease assay
Less than 10% of 1-μg of ΦX174 RFI is converted to RFII when the
DNA is incubated with 10 U of enzyme at 37℃ for
4hr in 50-μl reaction.
Ligation and recutting
More than 95% of DNA fragments (1 μg) digested with 10-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.