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237-PAd5.CMV-Cre-IRES-EGFP
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Cloning of Cre by subclone 241 to 242<br> Purpose: to clone 237-pAd5-Cre-GFP<br> Date: 2011-10-14 Notebook: luyingying Page: Clone ID:237 Experimenters: lu yingying Result <br> [[Image:237subclone10-13-2011.JPG]] band shows vector [[Image:237vector10-14-2011.JPG]] bands show cre<br> <br> Restart<br> Date: 2011-10-18 ---<br> Notebook: luyingying Page: Clone ID:237 Experimenters: lu yingying Result <br> [[Image:237 crePCR product10-18-2011.JPG]]<br> PCR product <br> 298 plasmid as template<br> primer:Lenti-Cre-F+Lenti-Cre-R<br> 25 cycles [[Image:237 vector+cre digest 10-19-2011.JPG]]<br> digest resulte:EcoR1+ Xho1 NEB enzyme 37 for 3 hours<br> 242 plasmid digest as vector<br> <br> Cloning of Cre by subclone 298 pTriEx-HTNC (Cre) to 242 pAd5-CMV-2-IRES-GFP<br> Purpose: to clone 237-pAd5-Cre-GFP Date: 2011-10-25 Notebook: luyingying Page: Clone ID:237 Experimenters: lu yingying Result <br> [[Image:470+359+237cloning 10-25-2011.JPG]]<br> band 4,5 show subclone result:EcoR1+ Xho1<br> band 4 show 242 pAd5-CMV-2-IRES-GFP digest result<br> band 5 show 298 pTriEx-HTNC (Cre) digest result<br> [[Image:237 digest detect10-27-2011.JPG]]<br> band 1-6 show digest detect result:EcoR1+ Xho1<br> <br> Date: 2011-11-10 --- Notebook: luyingying Page: Clone ID:237 Experimenters: lu yingying Result <br> [[Image:237 crePCR product10-18-2011.JPG]] PCR amplify His-Tat-NLS-Cre(298) as temlate, using Cre-EcoR1-F B1 + Cre-BglII-R B2<br> <br> [[Image:237+469 digest11-9-2011.JPG]] PCR product with EcoR1 + BglII, digest pAd5-CMV-IRES-GFP with BamH1 + EcoR1<br> band 1 show insert, band 2 show pAd5-CMV-IRES-GFP(242) vector digest<br> [[Image:237+469+493 digest detect 11-14-2011.JPG]] digest detect:EcoR1+ Not1<br> <br> Date: 2011-11-16 --- Notebook: luyingying Page: Clone ID:237 Experimenters: lu yingying Result <br> [[Image:237-PCR 11-16-2011.JPG]]<br> PCR amplify His-Tat-NLS-Cre(298) as temlate, using Cre-EcoR1-F2 B1 + Cre-BglII-R B2<br> [[Image:237 insert+vector11-17-2011.JPG]]<br> PCR product with EcoR1 + BglII, digest pAd5-CMV-IRES-GFP with BamH1 + EcoR1<br> [[Image:237 digest detect11-21-2011.JPG]] digest test:EcoR1+Not1 <br> <br> [[Category:Cloning]]
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