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2642 - Revision history
2026-10-18T13:20:38Z
Revision history for this page on the wiki
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Paul.xu at 01:22, 3 January 2020
2020-01-03T01:22:55Z
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<p><b>New page</b></p><div>[[File:2642 2643 2644 2598 2603.JPG|RTENOTITLE]]<br />
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[[File:2603 2642.JPG|RTENOTITLE]]<br />
<br />
both 2642 #2, #3 no priming<br />
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[[File:2603 digestion.png|RTENOTITLE]]<br />
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Expression screening using 10 ul magnetic beads from expedeon&nbsp;Amintra® Nickel Magnetic Affinity Resin<br />
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[[File:Cre375 2603 2642 2643 2644.jpg|RTENOTITLE]]<br />
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{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| Lane<br />
| ID<br />
| Name<br/><br />
| Host<br />
| MW<br />
| Induction<br />
| Note<br />
| Expression<br />
|-<br />
| 1<br />
| 375<br />
| pET28-Cre<br/><br />
| Nico21(DE3)pLysN,&nbsp;<br />
| 10.30 / 42.8<br/><br />
| 1 mM IPTG, 18 oC<br />
| grown on LB/KC, most cells are lysed<br/><br />
| y<br />
|-<br />
| 2<br />
| 375<br />
| pET28-Cre<br/><br />
| Rosetta(DE3)<br />
| 10.30 / 42.8<br/><br />
| 1 mM IPTG<br/><br />
| looks like contaminated, can not lyse cells<br />
| trace<br />
|-<br />
| 3<br />
| 375<br />
| pET28-Cre<br />
| <br />
Nico21(DE3)pLysN<br />
<br />
<br />
<br />
| 10.30 / 42.8<br/><br />
| 1 mM IPTG<br/><br />
| grown on LB/K<br/><br />
| y<br />
|-<br />
| 4<br />
| 2603<br />
| pL6H-sso7d-SD-Taq<br/><br />
| Nico21(DE3)pLysN<br/><br />
| 6.10 / 104.7<br/><br />
| 1 mM IPTG<br/><br />
| <br/><br />
| y<br />
|-<br />
| 5<br />
| 2642<br />
| pET29b-Flp<br/><br />
| Nico21(DE3)pLysN<br/><br />
| 9.76 / 52.5<br/><br />
| 1 mM IPTG<br/><br />
| most cells are lysed<br />
| n<br />
|-<br />
| M<br />
| <br/><br />
| <br/><br />
| pageruler plus<br />
| <br/><br />
| <br/><br />
| <br/><br />
| <br/><br />
|-<br />
| 6<br />
| 2643<br />
| pET29b-Int-C3<br/><br />
| Nico21(DE3)pLysN<br/><br />
| 9.69 / 41.4<br/><br />
| 1 mM IPTG<br/><br />
| <br/><br />
| maybe<br />
|-<br />
| 7<br />
| 2643<br/><br />
| pET29b-Int-C3<br/><br />
| Nico21(DE3)pLysN<br/><br />
| 9.69 / 41.4<br/><br />
| 1 mM IPTG<br/><br />
| <br/><br />
| maybe<br />
|-<br />
| 8<br />
| 2644<br />
| pET29b-scIHF2<br/><br />
| Nico21(DE3)pLysN<br/><br />
| 9.60 / 22.8<br/><br />
| 1 mM IPTG<br/><br />
| <br/><br />
| &nbsp;?<br />
|-<br />
| 9<br />
| 1010<br />
| Cre<br />
| Nico21(DE3)pLysN<br/><br />
| <br/><br />
| <br/><br />
| Lot&nbsp;174-1010<br />
| <br/><br />
|-<br />
| 10<br />
| 1186<br />
| MBP or MBP-Xis<br/><br />
| <br/><br />
| <br/><br />
| <br/><br />
| <br />
| <br/><br />
|}<br />
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1/2/2020, small scale expression screening using magnetic beads, 15 ml culture medium, induction at 37 oC<br />
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{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| Lane<br />
| ID<br />
| Sample<br />
| Competent Cells<br />
| Induction<br />
|-<br />
| 1<br />
| 2642<br />
| Soluble<br />
| BL21-AI<br />
| 0.2% arabinose 4 hours<br />
|-<br />
| 2<br />
| 2642<br/><br />
| Inclusion body<br />
| BL21-AI<br/><br />
| 0.2% arabinose 4 hours<br/><br />
|-<br />
| 3<br />
| 2642<br/><br />
| Soluble<br/><br />
| Nico21(DE3)<br />
| 0.5M IPTG 4 hours<br/><br />
|-<br />
| 4<br />
| 2642<br/><br />
| Inclusion body<br/><br />
| Nico21(DE3)<br/><br />
| 0.5M IPTG 4 hours<br/><br />
|-<br />
| 5<br />
| 2643<br />
| soluble<br />
| Nico21(DE3)<br />
| 0.5M IPTG 4 hours<br/><br />
|-<br />
| 6<br />
| 2644<br />
| soluble<br/><br />
| Nico21(DE3)<br/><br />
| 0.5M IPTG 4 hours<br/><br />
|-<br />
| M<br />
| PageRuler Plus<br />
| <br/><br />
| <br/><br />
| <br/><br />
|}<br />
<br />
[[Category:Cloning]]<br />
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[[File:2642 2643 2644.jpg]]</div>
Paul.xu