https://eln.excellgen.com/index.php?action=history&feed=atom&title=1708_1823_2116_2133_2115_2108 1708 1823 2116 2133 2115 2108 - Revision history 2026-10-18T05:50:27Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1708_1823_2116_2133_2115_2108&diff=16632&oldid=prev Paul.xu at 18:47, 19 November 2017 2017-11-19T18:47:27Z <p></p> <p><b>New page</b></p><div>Expression screening, 4/2/2014, by Shanyi<br /> <br /> PageRuler Plus 10, 15, 25(red), 35, 55, 70(red), 100, 130, 250<br /> <br /> 0.5 mM IPTG induction o/n at RT, 5 ml culture<br /> <br /> Lyse cell by freeze-thawing, 3 cycles, using Buffer A, spin--&gt; Supernatant (S)<br /> <br /> Add lysis buffer containing 4M urea, 1% SDS in Buffer A to pellet, solubilize and spin--&gt;P<br /> <br /> [[File:1708 1823 2116 2133 exp screening 4 3 14.jpg|RTENOTITLE]]<br /> <br /> {| width=&quot;554&quot; border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | Lane&lt;br/&gt;<br /> | Clone ID&lt;br/&gt;<br /> | Name&lt;br/&gt;<br /> | Competent Cells&lt;br/&gt;<br /> | Sample&lt;br/&gt;<br /> | Expression&lt;br/&gt;<br /> | Note&lt;br/&gt;<br /> |-<br /> | 1&lt;br/&gt;<br /> | 1708&lt;br/&gt;<br /> | pT7A3-HTNCre&lt;br/&gt;<br /> | Tuner(DE3)&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 2&lt;br/&gt;<br /> | 1708&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | IB?&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | &#039;&#039;&#039;3&#039;&#039;&#039;&lt;br/&gt;<br /> | 1823&lt;br/&gt;<br /> | pCold-HTNCre&lt;br/&gt;<br /> | Nico(DE3)pLysN&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &#039;&#039;&#039;Soluble&#039;&#039;&#039;&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | M&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | Fermentas&lt;br/&gt;<br /> | PageRuler Plus&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 4&lt;br/&gt;<br /> | 1823&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 5&lt;br/&gt;<br /> | 1823&lt;br/&gt;<br /> | pCold-HTNCre&lt;br/&gt;<br /> | Tuner(DE3)&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 6&lt;br/&gt;<br /> | 1823&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | IB?&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 7&lt;br/&gt;<br /> | 2116&lt;br/&gt;<br /> | pT7A3-Penetratin-Cre&lt;br/&gt;<br /> | Tuner(DE3)&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | No&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 8&lt;br/&gt;<br /> | 2116&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 9&lt;br/&gt;<br /> | 2133&lt;br/&gt;<br /> | pT7A3-ILK&lt;br/&gt;<br /> | BL21AI-pRare2LysS&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 10&lt;br/&gt;<br /> | 2133&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | low&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 11&lt;br/&gt;<br /> | 2133&lt;br/&gt;<br /> | pT7A3-ILK&lt;br/&gt;<br /> | Tuner(DE3)-pRare2LysS&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &#039;&#039;&#039;Soluble&#039;&#039;&#039;&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |}<br /> <br /> <br /> <br /> [[File:2133 2115 2108 exp screening 4 3 14.jpg|RTENOTITLE]]<br /> <br /> <br /> <br /> {| width=&quot;554&quot; border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | Lane&lt;br/&gt;<br /> | Clone ID&lt;br/&gt;<br /> | Name&lt;br/&gt;<br /> | Competent Cells&lt;br/&gt;<br /> | Sample&lt;br/&gt;<br /> | Expression&lt;br/&gt;<br /> | Note&lt;br/&gt;<br /> |-<br /> | 1&lt;br/&gt;<br /> | 2133&lt;br/&gt;<br /> | pT7A3-ILK&lt;br/&gt;<br /> | Tuner(DE3)-pRare2LysS&lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | some&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 2&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | pT7A3-iFLP&lt;br/&gt;<br /> | Tuner(DE3)&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | &#039;&#039;&#039;3&#039;&#039;&#039;&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | IB?&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 4&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | Nico(DE3)pRarepLysS&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 5&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | IB?, low&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 6&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | Nico(DE3)pLysN&lt;br/&gt;<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | cell death&lt;br/&gt;<br /> |-<br /> | M&lt;br/&gt;<br /> | Fermentas&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | PageRuler Plus&lt;br/&gt;<br /> |-<br /> | 7&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | cell death&lt;br/&gt;<br /> |-<br /> | 8&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | BL21(DE3)pRareLysS<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 9&lt;br/&gt;<br /> | 2115&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | IB?&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 10&lt;br/&gt;<br /> | 2108&lt;br/&gt;<br /> | pCold-Tat-NLS-iFLP-6H&lt;br/&gt;<br /> | Tuner(DE3)<br /> | S&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 11&lt;br/&gt;<br /> | 2108&lt;br/&gt;<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> | P&lt;br/&gt;<br /> | IB?&lt;br/&gt;<br /> | &lt;br/&gt;<br /> |}<br /> <br /> <br /> [[Category:Protein Purification]]</div> Paul.xu