http://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&feed=atom&action=historyTeam:Caltech/Protocols/LacZ Assay - Revision history2024-03-29T06:43:58ZRevision history for this page on the wikiMediaWiki 1.16.5http://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=99176&oldid=prevCbeisel at 00:43, 30 October 20082008-10-30T00:43:35Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and grow for 3 hours at 37˚ and 225 RPM. The OD should be between 0.09 and 0.20 for 200 <del class="diffchange diffchange-inline">ul </del>in a 96-well plate measured in a plate reader (Spectramax M2).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and grow for 3 hours at 37˚ and 225 RPM. The OD should be between 0.09 and 0.20 for 200 <ins class="diffchange diffchange-inline">µl </ins>in a 96-well plate measured in a plate reader (Spectramax M2).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 <del class="diffchange diffchange-inline">ul</del>/mL ß-mercaptoethanol.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 <ins class="diffchange diffchange-inline">µl</ins>/mL ß-mercaptoethanol.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 100 µL of lysed cells with 600 µL substrate solution.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 100 µL of lysed cells with 600 µL substrate solution.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Substrate Solution: 60 mM Na2PO4, 40 mM NaH<sub>2</sub>HPO<sub>4</sub>, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Substrate Solution: 60 mM Na2PO4, 40 mM NaH<sub>2</sub>HPO<sub>4</sub>, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># When a faint yellow color is observed, add <del class="diffchange diffchange-inline">300ul </del>1M sodium carbonate to stop the <del class="diffchange diffchange-inline">solution</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># When a faint yellow color is observed, add <ins class="diffchange diffchange-inline">300 µl </ins>1M sodium carbonate to stop the <ins class="diffchange diffchange-inline">reaction</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Miller units are calculated using the following equation: MU = 1000 (ABS<sub>420</sub> / (0.02 * t * ABS<sub>600</sub>)).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Miller units are calculated using the following equation: MU = 1000 (ABS<sub>420</sub> / (0.02 * t * ABS<sub>600</sub>)).</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>}}</div></td></tr>
</table>Cbeiselhttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=99142&oldid=prevCbeisel at 00:42, 30 October 20082008-10-30T00:42:18Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and grow for 3 hours at 37˚ and 225 RPM <del class="diffchange diffchange-inline">to an </del>OD between 0.09 and 0.20 <del class="diffchange diffchange-inline">as assessed by </del>plate reader (Spectramax M2).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and grow for 3 hours at 37˚ and 225 RPM<ins class="diffchange diffchange-inline">. The </ins>OD <ins class="diffchange diffchange-inline">should be </ins>between 0.09 and 0.20 <ins class="diffchange diffchange-inline">for 200 ul in a 96-well plate measured in a </ins>plate reader (Spectramax M2).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</div></td></tr>
</table>Cbeiselhttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=99116&oldid=prevCbeisel at 00:41, 30 October 20082008-10-30T00:41:12Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and grow for 3 hours at 37˚ and 225 RPM to an OD between 0.09 and 0.20 as assessed by plate reader ().</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and grow for 3 hours at 37˚ and 225 RPM to an OD between 0.09 and 0.20 as assessed by plate reader (<ins class="diffchange diffchange-inline">Spectramax M2</ins>).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</div></td></tr>
</table>Cbeiselhttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=99109&oldid=prevCbeisel at 00:40, 30 October 20082008-10-30T00:40:47Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Grow cells overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and <del class="diffchange diffchange-inline">grown </del>for 3 hours at 37˚ and 225 RPM<del class="diffchange diffchange-inline">.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># In the morning, back-dilute overnight cultures 1/100 (50 µL into 5mL) and <ins class="diffchange diffchange-inline">grow </ins>for 3 hours at 37˚ and 225 RPM to <ins class="diffchange diffchange-inline">an </ins>OD between 0.09 and 0.20 <ins class="diffchange diffchange-inline">as assessed by plate reader ()</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">#* Regrow cells </del>to OD <del class="diffchange diffchange-inline">values </del>between 0.09 and 0.20. </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;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Combine 20 µL of cell culture and 380 µL permeabilization solution, mix, and place at 37˚C for 10 minutes.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#* Permeabilization Solution: 100mM Na<sub>2</sub>HPO<sub>4</sub>, 20mM KCl, 2 mM MgSO<sub>4</sub>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</div></td></tr>
</table>Cbeiselhttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=58918&oldid=prevJkm at 16:16, 20 October 20082008-10-20T16:16:13Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">ß-galactosidase assays were performed as follows. Cells were grown </del>overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. <del class="diffchange diffchange-inline">Overnight cultures were </del>back-<del class="diffchange diffchange-inline">diluted </del>1/100 (50 µL into 5mL) and grown for 3 hours at 37˚ and 225 RPM. <del class="diffchange diffchange-inline">OD600 values were taken on the spectrophotometer. Cells were used at </del>OD values between 0.09 and 0.20. 20 µL of <del class="diffchange diffchange-inline">our </del>cell culture and 380 µL permeabilization solution <del class="diffchange diffchange-inline">(</del>100mM <del class="diffchange diffchange-inline">Na2HPO4</del>, 20mM KCl, 2 mM <del class="diffchange diffchange-inline">MgSO4</del>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol<del class="diffchange diffchange-inline">) were mixed and placed at 37˚C for 10 minutes</del>. 100 µL of lysed cells <del class="diffchange diffchange-inline">were then mixed </del>with substrate solution <del class="diffchange diffchange-inline">(</del>60 mM Na2PO4, 40 mM <del class="diffchange diffchange-inline">NaH2PO4</del>, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol<del class="diffchange diffchange-inline">)</del>. When a faint yellow color <del class="diffchange diffchange-inline">was </del>observed, 300ul 1M sodium carbonate <del class="diffchange diffchange-inline">was added </del>to stop the solution. <del class="diffchange diffchange-inline"><BR></del>Miller units <del class="diffchange diffchange-inline">were </del>calculated using the following equation: </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Grow cells </ins>overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>1000 (<del class="diffchange diffchange-inline">ABS420 </del>/ (0.02 <del class="diffchange diffchange-inline">mL </del>* t * <del class="diffchange diffchange-inline">ABS600</del>).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># In the morning, </ins>back-<ins class="diffchange diffchange-inline">dilute overnight cultures </ins>1/100 (50 µL into 5mL) and grown for 3 hours at 37˚ and 225 RPM.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">#* Regrow cells to </ins>OD values between 0.09 and 0.20. </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"># Combine </ins>20 µL of cell culture and 380 µL permeabilization solution<ins class="diffchange diffchange-inline">, mix, and place at 37˚C for 10 minutes.</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">#* Permeabilization Solution: </ins>100mM <ins class="diffchange diffchange-inline">Na<sub>2</sub>HPO<sub>4</sub></ins>, 20mM KCl, 2 mM <ins class="diffchange diffchange-inline">MgSO<sub>4</sub></ins>, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol.</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"># Combine </ins>100 µL of lysed cells with <ins class="diffchange diffchange-inline">600 µL </ins>substrate solution<ins class="diffchange diffchange-inline">.</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">#* Substrate Solution: </ins>60 mM Na2PO4, 40 mM <ins class="diffchange diffchange-inline">NaH<sub>2</sub>HPO<sub>4</sub></ins>, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol.</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>When a faint yellow color <ins class="diffchange diffchange-inline">is </ins>observed, <ins class="diffchange diffchange-inline">add </ins>300ul 1M sodium carbonate to stop the solution.</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>Miller units <ins class="diffchange diffchange-inline">are </ins>calculated using the following equation: <ins class="diffchange diffchange-inline">MU = </ins>1000 (<ins class="diffchange diffchange-inline">ABS<sub>420</sub> </ins>/ (0.02 * t * <ins class="diffchange diffchange-inline">ABS<sub>600</sub>)</ins>).</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>}}</div></td></tr>
</table>Jkmhttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=58646&oldid=prevRobertOvadia at 05:46, 20 October 20082008-10-20T05:46:25Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>ß-galactosidase assays were performed as follows. Cells were grown overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. Overnight cultures were back-diluted 1/100 (50 µL into 5mL) and grown for 3 hours at 37˚ and 225 RPM. OD600 values were taken on the spectrophotometer. Cells were used at OD values between 0.09 and 0.20. 20 µL of our cell culture and 380 µL permeabilization solution (100mM Na2HPO4, 20mM KCl, 2 mM MgSO4, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol) were mixed and placed at 37˚C for 10 minutes. 100 µL of lysed cells were then mixed with substrate solution (60 mM Na2PO4, 40 mM NaH2PO4, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol). When a faint yellow color was observed, 300ul 1M sodium carbonate was added to stop the solution. Miller units were calculated using the following equation: </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>ß-galactosidase assays were performed as follows. Cells were grown overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. Overnight cultures were back-diluted 1/100 (50 µL into 5mL) and grown for 3 hours at 37˚ and 225 RPM. OD600 values were taken on the spectrophotometer. Cells were used at OD values between 0.09 and 0.20. 20 µL of our cell culture and 380 µL permeabilization solution (100mM Na2HPO4, 20mM KCl, 2 mM MgSO4, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol) were mixed and placed at 37˚C for 10 minutes. 100 µL of lysed cells were then mixed with substrate solution (60 mM Na2PO4, 40 mM NaH2PO4, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol). When a faint yellow color was observed, 300ul 1M sodium carbonate was added to stop the solution. <ins class="diffchange diffchange-inline"><BR></ins>Miller units were calculated using the following equation: </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>1000 (ABS420 / (0.02 mL * t * ABS600).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>1000 (ABS420 / (0.02 mL * t * ABS600).</div></td></tr>
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</table>RobertOvadiahttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=58645&oldid=prevRobertOvadia at 05:45, 20 October 20082008-10-20T05:45:55Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Insert protocol here</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">ß-galactosidase assays were performed as follows. Cells were grown overnight in 5 mL culture of LB Media with proper antibiotic at 37˚C and 225 RPM. Overnight cultures were back-diluted 1/100 (50 µL into 5mL) and grown for 3 hours at 37˚ and 225 RPM. OD600 values were taken on the spectrophotometer. Cells were used at OD values between 0.09 and 0.20. 20 µL of our cell culture and 380 µL permeabilization solution (100mM Na2HPO4, 20mM KCl, 2 mM MgSO4, 0.6 mg/mL CTAB, 0.4 mg/mL sodium deoxycholate, and 5.4 ul/mL ß-mercaptoethanol) were mixed and placed at 37˚C for 10 minutes. 100 µL of lysed cells were then mixed with substrate solution (60 mM Na2PO4, 40 mM NaH2PO4, 1 mg/mL ONPG, and 5.4 µL/mL ß-mercaptoethanol). When a faint yellow color was observed, 300ul 1M sodium carbonate was added to stop the solution. Miller units were calculated using the following equation: </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">1000 (ABS420 / (0.02 mL * t * ABS600).</ins></div></td></tr>
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</table>RobertOvadiahttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=43537&oldid=prevJkm at 20:38, 8 September 20082008-09-08T20:38:15Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><font face="verdana" style="color:#<del class="diffchange diffchange-inline">BB4400</del>">LacZ Assay</font></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><font face="verdana" style="color:#<ins class="diffchange diffchange-inline">CC3300</ins>">LacZ Assay</font></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>__NOTOC__</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>__NOTOC__</div></td></tr>
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</table>Jkmhttp://2008.igem.org/wiki/index.php?title=Team:Caltech/Protocols/LacZ_Assay&diff=43510&oldid=prevJkm: New page: {{Caltech_iGEM_08| Content= <div style="font-size:18pt;"> <font face="verdana" style="color:#BB4400">LacZ Assay</font></div> __NOTOC__ <br> Insert protocol here }}2008-09-08T18:37:51Z<p>New page: {{Caltech_iGEM_08| Content= <div style="font-size:18pt;"> <font face="verdana" style="color:#BB4400">LacZ Assay</font></div> __NOTOC__ <br> Insert protocol here }}</p>
<p><b>New page</b></p><div>{{Caltech_iGEM_08|<br />
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