"
Team:Oxford/protocols/DpnI Digest
From 2014.igem.org
(Difference between revisions)
(Created page with "__NOTOC__ {{:Team:Oxford/templates/header}} <div class="abcdefg"> <div class="container cf row"> <h1>DpnI Digest</h1> <p>DpnI is a restriction enzyme that cleaves methylated DNA...") |
|||
| Line 5: | Line 5: | ||
<h1>DpnI Digest</h1> | <h1>DpnI Digest</h1> | ||
| - | <p>DpnI is a restriction enzyme that cleaves methylated DNA | + | <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> |
<!--this will number your steps 1,2,3,etc--> | <!--this will number your steps 1,2,3,etc--> | ||
Revision as of 14:53, 21 July 2014
DpnI Digest
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).
