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Bgl I
Recognition & cleavage sequence
Source: Bacillus
globigii
Reaction
conditions
1X
EzBuffer III 37℃
1X
FastCut Buffer, 37℃
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 37℃ for 1 hr in 50-μl reaction mixtures.
Storage
20
mM Tris-HCl (pH 7.5 @ 25℃), 200 mM
NaCl, 0.1 mM EDTA, 10 mM 2-mercaptoethanol, 200 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
Bgl
I can be inactivated at 65℃ for 20 min.
Methylation
sensitivity
dam methylation:
Not sensitive
dcm methylation:
Not sensitive
CpG methylation:
Conditionally sensitive
Prolonged
incubation
A
minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 0.13
U.
Relative
activity in EzBuffers
EzBuffer
I: 75%
EzBuffer
II: 75%
EzBuffer
III: 100%
EzBuffer
IV: 50%
FastCut
Buffer: 100%
Note
Five-fold
higher activity can be obtained at pH 9.5 than pH 7.5. Cleavage of supercoiled
DNA requires more (10 fold) enzyme. The sequences of sticky ends produced by
this enzyme are unique to each site, thus, this enzyme can be conveniently used
for unique cloning strategies; replacement of a wild-type fragment with a
mutant one in a plasmid. Cleavage of mammalian genomic DNA can be blocked by
CpG methylation that partially overlaps its recognition sequence. It is
sensitive to impurities in DNA and buffer condition.
● 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 5% of 1-μg of ΦX174 RFI is
converted to RFII when the DNA is incubated with 50 U of enzyme at 37℃ for 4hr in 50-μl reaction.
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.