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1745-p8HIC-sfGFP - Revision history
2026-10-18T11:59:24Z
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Paul.xu at 18:25, 22 May 2013
2013-05-22T18:25:50Z
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<p><b>New page</b></p><div>== '''Cloning of 1745-p8HIC-sfGFP''' ==<br />
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'''Purpose:''' Cloning of 1745-p8HIC-sfGFP <br />
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'''Date: '''2013-5-8---2013-5-16 <br />
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'''Notebook:0''' <br />
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'''Page:''' <br />
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'''Clone ID:1745''' <br />
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'''Experimenters:''' yingying <br />
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== '''Results''' ==<br />
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=== cloning ===<br />
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[[Image:553+559+1724+1741+1745clone 5-8-2013.JPG]] <br />
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=== Colony Screening ===<br />
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Date: 2013-5-16 <br />
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Make miniprep and digest with Nhe1 + Xho1 <br />
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[[Image:1741+1745 digest 5-16-2013.JPG]]<br> <br />
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<br> <br />
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== '''Result intepretation/Summary''' ==<br />
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<br>== '''Conclusion''' ==<br><br> <br />
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== protein screening ==<br />
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= 2013-5-21<br> =<br />
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[[Image:1724+1729+1731+1741+1742+1743+1744+1745+1748+1749 protein screen 5-21-2013.jpg]]<br> <br />
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Ni beads <br />
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{| cellspacing="1" cellpadding="1" border="1" width="254"<br />
|-<br />
| lane<br> <br />
| clone ID<br> <br />
| host<br> <br />
| induction<br> <br />
| purification<br> <br />
| MW<br><br />
|-<br />
| 1<br> <br />
| 1694,pT7A3-wtSM2<br> <br />
| BL21(DE3)pRare2pLysS<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| <br />
CBD beads, <br />
<br />
wrong buffer,need to do again <br />
<br />
| 33KD<br><br />
|-<br />
| 2<br> <br />
| 1724,pT7A3-6His-sc-VVCap<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 70KD<br><br />
|-<br />
| 3<br> <br />
| 1729,pCold-TF-(E3C)-3C<br> <br />
| BL21(DE3)pRare2pLysS<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 51KD+21KD<br><br />
|-<br />
| 4<br> <br />
| 1731,pT7A3-TurboEnterokinase<br> <br />
| BL21(DE3)pRare2pLysS<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 27KD<br><br />
|-<br />
| 5<br> <br />
| 1741,pIC-sfGFP<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| no his tag,supernatant run gel<br> <br />
| 31KD<br><br />
|-<br />
| 6<br> <br />
| 1742,p8H-ELK-IN<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 15KD<br><br />
|-<br />
| 7<br> <br />
| 1743,pIC<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 4KD<br><br />
|-<br />
| 8<br> <br />
| 1744,p8HIC<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 5KD<br><br />
|-<br />
| 9<br> <br />
| 1745,p8HIC-sfGFP<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads,lysis supernatant is green <br> <br />
| 15KD<br><br />
|-<br />
| 10<br> <br />
| 1748,pCold-8His-MBP-NRF2-6His<br> <br />
| BL21(DE3)pRare2pLysS<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br><br />
|-<br />
| 11<br> <br />
| 1749,p7H-ELK-IN<br> <br />
| NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 15KD<br><br />
|}<br />
<br />
<br> <br />
<br />
[[Category:Cloning]]</div>
Paul.xu