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BglI
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BglI is a restriction enzyme which recognizes the following sequence:
GCCN NNN▼NGGC
CGGN▲NNN NCCG |
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Concentration
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10u/μl
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Reaction condition |
Recommended Buffer: Buffer O+
50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. |
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Incubation temperature: 37°C |
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Thermal inactivation: 65°C |
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Source
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Bacillus globigii.
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Frequency of Cutting
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| λ | Ad-2 | ΦX174 | pUC18 | M13mp18 | pBR322 |
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| 29 | 20 | 0 | 2 | 1 | 3 |
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Storage
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-20°C
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Storage
buffer
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10mM Tris-HCl (pH 7.5
at 25°C), 300mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml
BSA and 50% glycerol
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Ligation
and Recleavage
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After 50-fold overdigestion
with BglI, approximately more than 90% of the DNA fragments
can be ligated and recut
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Methylation
Effect
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BglI does not cut Gm5CCN5GGC and
GCm4CN5GCC,
but cuts GCm5CN5CN5GGC
The hemi-methylated sequence GCCN5GGm5C is cleaved by
BglI at a significantly slower rate
Cleavage is blocked when both strands are methylated as
shown
5'...GCCN5GGm5C...3'
3'... m5CGGN5CC G...5'
Overlapping GC methylation may influence DNA cleavage |
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