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RsaI
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RsaI is a restriction enzyme which recognizes the following sequence:
GT▼AC
CA▲TG |
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Concentration
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10u/μl
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Reaction condition |
Recommended Buffer: Y+/TANGO™
33mM Tris-acetate (pH 7.9), 10mM Mg-acetate, 66mM K-acetate
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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Rhodopseudomonas sphaeroides.
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Frequency of Cutting
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| λ | Ad-2 | ΦX174 | pUC18 | M13mp18 | pBR322 |
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| 113 | 83 | 11 | 3 | 19 | 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 7.4
at 25°C), 100mM KCl, 1mM DTT, 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 RsaI, approximately more than 90% of the DNA fragments
can be ligated and recut
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Methylation
Effect
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RsaI cuts neither GTm6AC nor GTAm4C
The hemi-methylated sequence GTAm5C is cleaved by RsaI
at a significantly slower rate
Cleavage is blocked when both strands are methylated
as shown
5'...GTAm5C...3'
3'...m5CATG...5'
Overlapping CG methylation may influence DNA cleavage |
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