http://2007.igem.org/wiki/index.php?title=Melbourne/Lab_Notebook_gv_F1&feed=atom&action=historyMelbourne/Lab Notebook gv F1 - Revision history2024-03-28T23:46:16ZRevision history for this page on the wikiMediaWiki 1.16.5http://2007.igem.org/wiki/index.php?title=Melbourne/Lab_Notebook_gv_F1&diff=41864&oldid=prevPhillipDodson at 13:50, 26 October 20072007-10-26T13:50:12Z<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 13:50, 26 October 2007</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 15:</td>
<td colspan="2" class="diff-lineno">Line 15:</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>==Example pNL29(orininal plasmid) and Tube(64) the IPTG promoted version of BBa_I750016==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Example pNL29(orininal plasmid) and Tube(64) the IPTG promoted version of BBa_I750016==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Image:Melbourne-float1 11am 18-10-2007.JPG</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">In the following series of photos the tubes are in pairs. The outer pairs are uninduced (no IPTG), the inner pairs are induced. The tubes left of centre are the original pNL29 plasmid in DH5a cells, the tubes on the right are 41C plasmids (promoted BBa_I750016) in XL10 gold. We believe the XL10-gold is permanenently inducing the expression even without IPTG, this seems to be confirmed by the latter experiment where the plasmid was retransformed into DH5alpha cell. </ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[Image:Melbourne-float1 6pm 17-10-2007.jpg|850px|thumb|6hrs (6pm 17-10-2007)]]</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[</ins>Image:Melbourne-float1 11am 18-10-2007.JPG<ins class="diffchange diffchange-inline">|850px|thumb|11hrs (11Am 18-10-2007)]]</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[Image:Melbourne-float1 6pm 20-10-2007.jpg|850px|thumb|78hrs (6pm 20-10-2007)]]</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[Image:Melbourne-float1 4pm 21-10-2007.jpg|850px|thumb|100hrs (4pm 21-10-2007)]]</ins></div></td></tr>
</table>PhillipDodsonhttp://2007.igem.org/wiki/index.php?title=Melbourne/Lab_Notebook_gv_F1&diff=41730&oldid=prevPhillipDodson at 13:29, 26 October 20072007-10-26T13:29:30Z<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 13:29, 26 October 2007</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 15:</td>
<td colspan="2" class="diff-lineno">Line 15:</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>==Example pNL29(orininal plasmid) and Tube(64) the IPTG promoted version of BBa_I750016==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Example pNL29(orininal plasmid) and Tube(64) the IPTG promoted version of BBa_I750016==</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Image:Melbourne-float1 11am 18-10-2007.JPG</ins></div></td></tr>
</table>PhillipDodsonhttp://2007.igem.org/wiki/index.php?title=Melbourne/Lab_Notebook_gv_F1&diff=41589&oldid=prevPhillipDodson at 13:08, 26 October 20072007-10-26T13:08:46Z<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 13:08, 26 October 2007</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>[[Melbourne/Lab GV Notebook| <Return to lab book summary>]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Melbourne/Lab GV Notebook| <Return to lab book summary>]]</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">==Testing Floatation==</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*[[Melbourne/Growing up cells|Growing up cells from glycerol stocks overnight in 5mL LB amp]]</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Prepare 250ml conical shaker flasks by cleaning and autoclave.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Add 50mL of LB Amp to each of two flasks.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Add 1M IPTG to one flask to make final concentration of 1mM IPTG, Lable with I.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Transfer 1mL of overnight culture to each flask.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Grow overnight at 37degC on a shaker.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Next morning transfer to 50ml falcon tubes.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Spin down the cells gently 25minutes at 1000rpm in Eppendorph 5810R centrifuge.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Poor off and discard the LB, maintaining the pellet and a residual ammount of LB 1ml about.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Replace LB with 50ml of 10g/L NaCl filtered through a 0.22um filter.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Resuspend the cells by shaking.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Leave at room temperature and observe the settling over the next week.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">==Example pNL29(orininal plasmid) and Tube(64) the IPTG promoted version of BBa_I750016==</ins></div></td></tr>
</table>PhillipDodsonhttp://2007.igem.org/wiki/index.php?title=Melbourne/Lab_Notebook_gv_F1&diff=41322&oldid=prevPhillipDodson at 12:32, 26 October 20072007-10-26T12:32:06Z<p></p>
<p><b>New page</b></p><div>[[Melbourne/Lab GV Notebook| <Return to lab book summary>]]</div>PhillipDodson