Team:Marburg:Project:Notebook:May
From 2014.igem.org
(Difference between revisions)
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<tr> | <tr> | ||
<th scope="col">Fragment</th> | <th scope="col">Fragment</th> | ||
- | <th scope="col">Concentration [ng/ | + | <th scope="col">Concentration [ng/µl]</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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</tr> | </tr> | ||
</table> | </table> | ||
- | <p>Incubation for 1, | + | <p>Incubation for 1,5 h at room temperature.</p> |
</div> | </div> | ||
</fieldset> | </fieldset> | ||
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</fieldset> | </fieldset> | ||
</div> | </div> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.24">14.24 Transformation of BL21 cells</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Transform E. coli BL21(DE3) with PheA-Arc1p-C-2x-pET28a</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>40 µL E. coli BL21(DE3) electrocompetent cells were transformed with 1 ng of the previously purified plasmid PheA-Arc1p-C-2x-pET28a from two different clones, incubated for 1 h at 37 °C and plated out on LB-Kan<sup>50</sup> plates that were incubated overnight at 37 °C.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
<!-- 13.05.14 --> | <!-- 13.05.14 --> | ||
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<a name="13.05.2014">13.05.2014</a> | <a name="13.05.2014">13.05.2014</a> | ||
</h2> | </h2> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.25">14.25 Overnight culture</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Inoculate Overnight culture of PheA-Arc1p-C-2x-pET28a</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>5 mL of LB-Kan<sup>50</sup> medium were inoculated with a clone from 14.24 bearing the PheA-Arc1p-C-2x-pET28a plasmid.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.33">15.33 Miniprep and Restriction of pMAD with BamHI and NnoI</a></legend> | <legend><a name="exp15.33">15.33 Miniprep and Restriction of pMAD with BamHI and NnoI</a></legend> | ||
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<a name="14.05.2014">14.05.2014</a> | <a name="14.05.2014">14.05.2014</a> | ||
</h2> | </h2> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.26">14.26 Test Expression</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Test the expression of PheA-Arc1p-C-2x</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>Using the preculture from 14.25 60 mL of LB-Kan<sup>50</sup> medium were inoculated with 600 µL of the preculture and incubated at 37 °C and 220 rpm until OD<sub>600</sub> reached 0.5. The culture was cooled down to 18 °C and the protein production was induced with 12 µL of 500 mM IPTG. Over night the culture was incubated at 18 °C and 220 rpm. A glycerolstock of the strain containing the PheA-Arc1p-C-2x-pET28a plasmid was stored at -80 °C.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.35">15.35 Inoculation Miniprep and Restriction of pMAD with BamHI and NcoI</a></legend> | <legend><a name="exp15.35">15.35 Inoculation Miniprep and Restriction of pMAD with BamHI and NcoI</a></legend> | ||
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</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.27">14.27 Purification of test expression</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Purify PheA-Arc1p-C-2x on a small scale</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>The cell culture from 14.26 was centrifuged (17000 rpm, 20 min, 4 °C) and the pellet resuspended in 3 mL buffer A. Lysozyme was added to a final concentration of 1 mg/mL and incubated on ice for 30 min. Afterwards the cells were lysed using ultra sound (2x 18 pulses, 30 s on ice inbetween). To clear the suspension of cell debris it was centrifuged (13000 rpm, 15 min, 4 °C). The supernatant was purified using QIAGEN Ni-NTA Spin Columns. An SDS-PAGE gel showed that the protein is produced and can be purified using Ni affinity chromatography.</p> | ||
+ | <p>60 mL LB-Kan<sup>50</sup> medium was inoculated from the glycerolstock prepared in 14.26 and incubated at 37 °C and 220 rpm over night.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.38">15.38 Transformation of Ligated pMAD and the Restricted hag/flank-Construct</a></legend> | <legend><a name="exp15.38">15.38 Transformation of Ligated pMAD and the Restricted hag/flank-Construct</a></legend> | ||
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</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.28">14.28 Expression of PheA-Arc1p-C-2x</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Express PheA-Arc1p-C-2x for further use</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>10 x 500 mL LB-Kan<sup>50</sup> medium were inoculated with 5 mL of the overnight culture prepared in 14.27 and incubated at 37 °C and 220 rpm until OD<sub>600</sub> reached 0.5. The flasks were cooled to 18 °C and protein production was induced by adding 100 µL 500 mM IPTG. The cultures were incubated over night at 18 °C and 220 rpm. Afterwards the cells were collected by centrifugation (6000 rpm, 20 min, 4 °C), resuspended in buffer A and stored at -20 °C until further use.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
</div> | </div> | ||
Line 1,939: | Line 2,020: | ||
</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.29">14.29 Purification of PheA-Arc1p-C-2x</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Purify PheA-Arc1p-C-2x in three steps</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>The resuspended cells from 14.28 were lysed using a french press. The lysate was incubated on ice with 5 µg/mL DNAseA and RNAseI for 10 min and centrifuged (27000 rpm, 45 min, 4 °C) to pellet cell debris. The supernatant was filtered (0.45 µm) and PheA-Arc1p-C-2x purified from the cleared solution by NiNTA affinity chromatography. The combined elution fractions containing crude PheA-Arc1p-C-2x were concentrated to a volume of 2 mL and further purified using an anion exchange column to get rid of possible tRNA contaminations. Finally the buffer was changed to dialysis buffer and the protein concentrated and stored at -80 °C in aliquods for further use.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
</div> | </div> | ||
Line 1,947: | Line 2,043: | ||
<a name="20.05.2014">20.05.2014</a> | <a name="20.05.2014">20.05.2014</a> | ||
</h2> | </h2> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.30">14.30 Creating PheA-Arc1p-C-1x</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Using Round-the-horn mutagenesis to create PheA-Arc1p-C-1x-pET28a</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | The PCR with 5' phosphorylated TG_1xGSSG FP und TG_0xGSSG RP should generate the 7509 bp fragment for ligation from the methylated template PheA-Arc1p-C-8x-pET28a from 14.5.</p> | ||
+ | <p>The used Primer concentration was 1 | ||
+ | µM, the concentration of the template was 1 ng/µL and we used a hot start to avoid unspecific annealing of primers.</p> | ||
+ | <table width="100%" border="1"> | ||
+ | <tr> | ||
+ | <th scope="col">Content</th> | ||
+ | <th scope="col">Volume [µL]</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">Water</th> | ||
+ | <td>29.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">5x GC Buffer</th> | ||
+ | <td>10</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">DMSO</th> | ||
+ | <td>2.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">TG_1xGSSG FP</th> | ||
+ | <td>2.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">TG_0xGSSG RP</th> | ||
+ | <td>2.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">PheA-Arc1p-C-8x-pET28a</th> | ||
+ | <td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">dNTPs</th> | ||
+ | <td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">Phusion-Polymerase</th> | ||
+ | <td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">Total Volume</th> | ||
+ | <td>50</td> | ||
+ | </tr> | ||
+ | </table><br /> | ||
+ | <table width="100%" border="1"> | ||
+ | <tr> | ||
+ | <th scope="col">Step</th> | ||
+ | <th scope="col">Temperature [°C]</th> | ||
+ | <th scope="col">Time [min:sec]</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">1</th> | ||
+ | <td>98</td> | ||
+ | <td>2:00</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">2</th> | ||
+ | <td>add Polymerase</td> | ||
+ | <td> </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">3</th> | ||
+ | <td>98</td> | ||
+ | <td>0:10</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">4</th> | ||
+ | <td>67.5</td> | ||
+ | <td>0:30</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">5</th> | ||
+ | <td>72</td> | ||
+ | <td>4:00</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">6</th> | ||
+ | <td>go to 3</td> | ||
+ | <td>32x</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">7</th> | ||
+ | <td>72</td> | ||
+ | <td>10:00</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">8</th> | ||
+ | <td>4</td> | ||
+ | <td>hold</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <p>The amplified fragment was purified on an agarose gel on which it showed the correct size. The extracted linear template was then digested to degrade the original unmutated plasmid using DpnI.</p> | ||
+ | <table width="100%" border="1"> | ||
+ | <tr> | ||
+ | <th scope="col">Content</th> | ||
+ | <th scope="col">Volume [µL]</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">10x CutSmart Buffer</th> | ||
+ | <td>3.6</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">Plasmid</th> | ||
+ | <td>30</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">DpnI</th> | ||
+ | <td>3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th scope="row">Total Volume</th> | ||
+ | <td>36.6</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <p>The reaction mixture was incubated at 37 °C and 300 rpm for 1 h and afterwards heated to 80 °C for 20 min.</p> | ||
+ | <p>In order to create the circular plasmid 100 ng of the linear template were mixed with 1 µL 10x T4-Ligasebuffer, 1 µL T4-Ligase and water to give a total volume of 10 µL. This mixture was incubated for 9 h at 16 °C. The resulting plasmid was dialysed with water to decrease the salt concentration for the following transformation of E. coli Top10 electrocompetent cells with the plasmid.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.43">15.43 PCR to Amplify Cup 1 from Yeast gDNA</a></legend> | <legend><a name="exp15.43">15.43 PCR to Amplify Cup 1 from Yeast gDNA</a></legend> | ||
Line 2,029: | Line 2,258: | ||
</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.31">14.31 Overnight culture</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Amplify Plasmid in an overnight culture</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>Six clones of the transformed Top10 cells from 14.30 were picked and used to inoculate overnight cultures (6 x 5 mL).</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.45">15.45 Gibson Assembly of Restricted pMad-fla and cup 1 and Transformation into <em>E.Coli</em> XL1-Blue</a></legend> | <legend><a name="exp15.45">15.45 Gibson Assembly of Restricted pMad-fla and cup 1 and Transformation into <em>E.Coli</em> XL1-Blue</a></legend> | ||
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</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.32">14.32 Preparation of plasmids</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Purify the amplified plasmids from overnight cultures</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>The plasmids were purified from the overnight cultures created in 14.31 using Sigma GenElute Plasmid Miniprep Kit following the included protocol. The plasmids were eluted using 45 µL water.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
</div> | </div> | ||
Line 2,273: | Line 2,536: | ||
</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.33">14.33 Transformation of BL21 cells</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Transform E. coli BL21(DE3) with PheA-Arc1p-C-1x-pET28a</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>40 µL E. coli BL21(DE3) electrocompetent cells were transformed with 1 ng of the previously purified plasmid PheA-Arc1p-C-1x-pET28a from two different clones, incubated for 1 h at 37 °C and plated out on LB-Kan<sup>50</sup> plates that were incubated overnight at 37 °C.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp13"> | <fieldset class="exp13"> | ||
<legend><a name="exp13.49">13.49 PlasmidPprep from Yeast and Transformation of <em>E. coli</em> DH5Alpha</a></legend> | <legend><a name="exp13.49">13.49 PlasmidPprep from Yeast and Transformation of <em>E. coli</em> DH5Alpha</a></legend> | ||
Line 2,343: | Line 2,623: | ||
<a name="27.05.2014">27.05.2014</a> | <a name="27.05.2014">27.05.2014</a> | ||
</h2> | </h2> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.34">14.34 Overnight culture</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Inoculate Overnight culture of PheA-Arc1p-C-1x-pET28a</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>5 mL of LB-Kan<sup>50</sup> medium were inoculated with a clone from 14.33 bearing the PheA-Arc1p-C-1x-pET28a plasmid.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.48">15.48 Restriction of pET24d-fla with SpeI and Gibson-Assembly with cup1-1</a></legend> | <legend><a name="exp15.48">15.48 Restriction of pET24d-fla with SpeI and Gibson-Assembly with cup1-1</a></legend> | ||
Line 2,630: | Line 2,927: | ||
<li>Resuspend pellets in 2ml TFB2 </li> | <li>Resuspend pellets in 2ml TFB2 </li> | ||
<li>Pipett 200µl aliquots in precooled 1,5 ml tubes</li> | <li>Pipett 200µl aliquots in precooled 1,5 ml tubes</li> | ||
- | <li>store | + | <li>store at -80µC </li> |
</ul> | </ul> | ||
</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.35">14.35 Test Expression</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Test the expression of PheA-Arc1p-C-1x</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>Using the preculture from 14.34 60 mL of LB-Kan<sup>50</sup> medium were inoculated with 600 µL of the preculture and incubated at 37 °C and 220 rpm until OD<sub>600</sub> reached 0.5. The culture was cooled down to 18 °C and the protein production was induced with 12 µL of 500 mM IPTG. Over night the culture was incubated at 18 °C and 220 rpm. A glycerolstock of the strain containing the PheA-Arc1p-C-1x-pET28a plasmid was stored at -80 °C.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp15"> | <fieldset class="exp15"> | ||
<legend><a name="exp15.49a">15.49 Colony-PCR with transformed <em>E. coli</em> XLI-Blue including pet24d-Fla-Cup1-1</a></legend> | <legend><a name="exp15.49a">15.49 Colony-PCR with transformed <em>E. coli</em> XLI-Blue including pet24d-Fla-Cup1-1</a></legend> | ||
Line 3,032: | Line 3,346: | ||
</div> | </div> | ||
</fieldset> | </fieldset> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.36">14.36 Purification of test expression</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Purify PheA-Arc1p-C-1x on a small scale</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>The cell culture from 14.35 was centrifuged (17000 rpm, 20 min, 4 °C) and the pellet resuspended in 3 mL buffer A. Lysozyme was added to a final concentration of 1 mg/mL and incubated on ice for 30 min. Afterwards the cells were lysed using ultra sound (2x 18 pulses, 30 s on ice inbetween). To clear the suspension of cell debris it was centrifuged (13000 rpm, 15 min, 4 °C). The supernatant was purified using QIAGEN Ni-NTA Spin Columns. An SDS-PAGE gel showed that the protein is produced and can be purified using Ni affinity chromatography.</p> | ||
+ | <p>60 mL LB-Kan<sup>50</sup> medium was inoculated from the glycerolstock prepared in 14.35 and incubated at 37 °C and 220 rpm over night.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp13"> | <fieldset class="exp13"> | ||
<legend><a name="exp13.53">15.53 Inoculation of Minipreps from Nose-Tag-Plasmids</a></legend> | <legend><a name="exp13.53">15.53 Inoculation of Minipreps from Nose-Tag-Plasmids</a></legend> | ||
Line 3,058: | Line 3,390: | ||
<a name="30.05.2014">30.05.2014</a> | <a name="30.05.2014">30.05.2014</a> | ||
</h2> | </h2> | ||
+ | |||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | <fieldset class="exp14"> | ||
+ | <legend><a name="exp14.37">14.37 Expression of PheA-Arc1p-C-1x</a></legend> | ||
+ | <div class="aim"> | ||
+ | <p>Aim: Express PheA-Arc1p-C-1x for further use</p> | ||
+ | </div> | ||
+ | <div class="exp-content"> | ||
+ | <p>10 x 500 mL LB-Kan<sup>50</sup> medium were inoculated with 5 mL of the overnight culture prepared in 14.36 and incubated at 37 °C and 220 rpm until OD<sub>600</sub> reached 0.5. The flasks were cooled to 18 °C and protein production was induced by adding 100 µL 500 mM IPTG. The cultures were incubated over night at 18 °C and 220 rpm. Afterwards the cells were collected by centrifugation (6000 rpm, 20 min, 4 °C), resuspended in buffer A and stored at -20 °C until further use.</p> | ||
+ | </div> | ||
+ | </fieldset> | ||
+ | |||
+ | <!-- ROMAN --> | ||
+ | |||
+ | |||
<fieldset class="exp18"> | <fieldset class="exp18"> | ||
<legend><a name="exp18.7">18.7 Miniprep of <em>E.Coli</em> XL1-Blue with pMAD and pMAD-fla</a></legend> | <legend><a name="exp18.7">18.7 Miniprep of <em>E.Coli</em> XL1-Blue with pMAD and pMAD-fla</a></legend> |
Revision as of 23:32, 15 October 2014
Notebook: May