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Team:Oxford/protocols/DpnI Digest
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<p>DpnI is a restriction enzyme that cleaves methylated DNA. Treating PCR products with DpnI will therefore remove any template DNA derived from the bacteria. All the newly PCR-synthesised DNA will be unmethylated and thus unaffected.</p> | <p>DpnI is a restriction enzyme that cleaves methylated DNA. Treating PCR products with DpnI will therefore remove any template DNA derived from the bacteria. All the newly PCR-synthesised DNA will be unmethylated and thus unaffected.</p> | ||
Latest revision as of 15:38, 21 July 2014
DpnI Digest
↩ Back to other protocols.
DpnI is a restriction enzyme that cleaves methylated DNA. Treating PCR products with DpnI will therefore remove any template DNA derived from the bacteria. All the newly PCR-synthesised DNA will be unmethylated and thus unaffected.
- Keeping the enzyme on ice, add 1 μl of DpnI to the PCR tube containing your sample. Gently invert a few times to mix contents.
- Incubate at in a 37⁰c water bath for 60 mins.
NOTE: The enzyme does not need to be removed, as in a conventional restriction enzyme digest, as it will be denatured by the SDS in the loading dye (if running on gel) or by the transformation conditions (if transforming straight into bacteria).
