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HincII
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HincII is a restriction enzyme which recognizes the following sequence:
GT(T/C)▼(A/G)AC
CA(A/G)▲(T/C)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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Haemophilus influenzae Rc.
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Frequency of Cutting
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| λ | Ad-2 | ΦX174 | pUC18 | M13mp18 | pBR322 |
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| 35 | 25 | 13 | 1 | 1 | 2 |
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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), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml
BSA and 50% glycerol
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Ligation
and Recleavage
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After 50-fold overdigestion
with HincII, approximately more than 90% of the DNA fragments
can be ligated and recut
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Methylation
Effect
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HincII (HindII) cuts neither GTPyPum6AC
nor GTPyPuAhm5C
GTm5CGAC and the hemi-methylated sequence GTPyPuAm5C are
cleaved by HincII (HindII) at a significantly slower rate.
Cleavage is blocked when both strands are methylated as
shown:
5'...GTPyPuAm5C...3'
3'...m5CAPuPyTG...5'
Overlapping CG methylation may influence DNA cleavage
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