http://www.openwetware.org/index.php?title=Corum:DNA_Digestion&action=history Revision history for this page on the wiki en MediaWiki 1.13.2 Fri, 24 May 2013 15:43:33 GMT http://www.openwetware.org/index.php?title=Corum:DNA_Digestion&diff=608807&oldid=prev <p>New page: '''SC 14:53, 19 June 2012 (EDT)''': ==Overview== Standard DNA digest protocol using [http://www.neb.com NEB] restriction enzymes. ==Materials== For a DNA digestion of total volume V (in...</p> <p><b>New page</b></p><div>'''SC 14:53, 19 June 2012 (EDT)''':<br /> <br /> ==Overview==<br /> Standard DNA digest protocol using [http://www.neb.com NEB] restriction enzymes.<br /> <br /> ==Materials==<br /> <br /> For a DNA digestion of total volume V (in μL):<br /> <br /> * ~0.8V DNA<br /> * 0.1V 10X digestion buffer (1, 2, 3, or 4; [http://www.neb.com NEB])<br /> * 0.1V 10mg/ml BSA<br /> * 1 μL restriction enzyme 1<br /> * 1 μL restriction enzyme 2 (optional, for double digestion)<br /> <br /> ==Procedure==<br /> # Find appropriate enzymes(s), associated buffer, reaction temperature from [http://www.neb.com NEB].<br /> # Calculate the total volume of the reaction by taking the DNA volume + 1(2) μL for a single (double) digestion reaction. Divide this number by 0.8 to get the total reaction volume, V.<br /> # Calculate the volume of 10mg/ml BSA and 10X digestion buffer, which are both 0.1V.<br /> # Add the reaction components together in order listed above. Mix gently and spin.<br /> # Incubate at reaction temperature for 3 hr to overnight.<br /> <br /> ==Notes==<br /> Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!<br /> #List troubleshooting tips here. <br /> #You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.<br /> #Anecdotal observations that might be of use to others can also be posted here. <br /> <br /> Please sign your name to your note by adding &lt;font face=&quot;courier&quot;&gt;&lt;nowiki&gt;'''*~~~~''':&lt;/nowiki&gt;&lt;/font&gt; to the beginning of your tip.<br /> <br /> ==References==<br /> '''Relevant papers, books, and websites'''<br /> &lt;!-- If this protocol has papers or books associated with it, list those references here.--&gt;<br /> &lt;!-- Try the [[Template:FormatRef|FormatRef template]]--&gt;<br /> #[http://www.neb.com New England Biolabs]<br /> <br /> ==Contact==<br /> * [http://openwetware.org/wiki/User:Sean_P_Corum Sean P Corum]<br /> <br /> or instead, [[Talk:{{PAGENAME}}|discuss this protocol]]. <br /> <br /> &lt;!-- You can tag this protocol with various categories. See the [[Categories]] page for more information. --&gt;<br /> <br /> [[Category:Protocol]]<br /> <br /> [[Category:DNA]]<br /> <br /> [[Category:In vitro]]<br /> <br /> [[Category:Digestion]]<br /> <br /> [[Category:Restriction Enzyme]]</div> Tue, 19 Jun 2012 23:55:51 GMT Sean P Corum http://www.openwetware.org/wiki/Talk:Corum:DNA_Digestion