http://www.openwetware.org/index.php?title=AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media&feed=atom&action=history 2013-05-22T02:21:23Z Revision history for this page on the wiki MediaWiki 1.13.2 http://www.openwetware.org/index.php?title=AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media&diff=529111&oldid=prev 2011-08-11T23:38:37Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 23:38, 11 August 2011</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 1:</td> <td colspan="2" class="diff-lineno">Line 1:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Description==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Description==</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Starter cultures enable us </del>to <del class="diffchange diffchange-inline">get </del>a <del class="diffchange diffchange-inline">jump-start on culturing </del>bacteria <del class="diffchange diffchange-inline">to log-phase while we are expressing proteins</del>. We will usually make 4 <del class="diffchange diffchange-inline">starter cultures for our expressions</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The expression culture will be able </ins>to <ins class="diffchange diffchange-inline">support </ins>a <ins class="diffchange diffchange-inline">large number of </ins>bacteria <ins class="diffchange diffchange-inline">as they express our protein of interest</ins>. We will usually make 4 <ins class="diffchange diffchange-inline">flasks-worth of expression culture</ins>.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Protocol==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Protocol==</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># Prepare [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] in a <del class="diffchange diffchange-inline">250mL Erlenmeyer </del>flask</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*This needs to be done the day before protein expression</ins></div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## <del class="diffchange diffchange-inline">0.875g </del>of LB</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># Prepare [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] in a <ins class="diffchange diffchange-inline">2.8L Fernbach </ins>flask</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## <del class="diffchange diffchange-inline">35mL </del>of water</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">25g </ins>of LB</div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">1L </ins>of water</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Cover the flask with foil</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Cover the flask with foil</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># Autoclave</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># Autoclave <ins class="diffchange diffchange-inline">(You can only fit two flasks into the autoclave at one time so you need to plan accordingly and perform two runs in the autoclave)</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Allow the flask to cool</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Allow the flask to cool</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># Add <del class="diffchange diffchange-inline">35uL </del>of 1000X Antibiotic</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># Add <ins class="diffchange diffchange-inline">1mL </ins>of 1000X Antibiotic</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"># Inoculate with the bacteria you are culturing using one of the two following methods</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">## Using sterile methods, retrieve a single colony from a petrie dish and drop it into the flask</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">## Using sterile methods, add 5uL of a glycerol stock to the flask</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"># Place the flask in a shaker/incubator and culture overnight at 37C and 200rpm</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Notes==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Notes==</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==References==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==References==</div></td></tr> <!-- diff generator: internal 2013-05-22 02:21:24 --> </table>

Matt Hartings http://www.openwetware.org/index.php?title=AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media&diff=529110&oldid=prev 2011-08-11T23:05:52Z

<p>New page: ==Description== Starter cultures enable us to get a jump-start on culturing bacteria to log-phase while we are expressing proteins. We will usually make 4 starter cultures for our expressi...</p> <p><b>New page</b></p><div>==Description==<br /> Starter cultures enable us to get a jump-start on culturing bacteria to log-phase while we are expressing proteins. We will usually make 4 starter cultures for our expressions.<br /> <br /> ==Protocol==<br /> # Prepare [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] in a 250mL Erlenmeyer flask<br /> ## 0.875g of LB<br /> ## 35mL of water<br /> # Cover the flask with foil<br /> # Autoclave<br /> # Allow the flask to cool<br /> # Add 35uL of 1000X Antibiotic<br /> # Inoculate with the bacteria you are culturing using one of the two following methods<br /> ## Using sterile methods, retrieve a single colony from a petrie dish and drop it into the flask<br /> ## Using sterile methods, add 5uL of a glycerol stock to the flask<br /> # Place the flask in a shaker/incubator and culture overnight at 37C and 200rpm<br /> <br /> ==Notes==<br /> <br /> ==References==</div>

Matt Hartings