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KpnI
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KpnI is a restriction enzyme which recognizes the following sequence:
5'...AG▼CT...3'
3'...TC▲GA…5'
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Concentration
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10u/μl
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Reaction condition |
Recommended Buffer: Buffer KpnI+
10mM Tris-HCl (pH 7.5), 10mM MgCl2, 0.02% Triton X-100 and 0.1mg/ml BSA. |
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Incubation temperature: 37°C |
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Thermal inactivation: 80°C |
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Source
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Recombinant E. coli strain.
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Frequency of Cutting
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| λ | Ad-2 | ΦX174 | pUC18 | M13mp18 | pBR322 |
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| 2 | 8 | 0 | 1 | 1 | 0 |
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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), 50mM KCl, 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 KpnI, approximately more than 90% of the DNA fragments
can be ligated and recut.
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Methylation
Effect
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KpnI cuts neither GGTm6ACC,
GGTAm4CC, GGTAm5Cm5C nor GGTACm4C, but cuts GGTAm5CC and
GGTACm5C.
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Notes |
Star activity may be observed with glycerol concentrations >12% or with enzyme:DNA ratios >25u/μg. |
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