237-pAd5-Cre-GFP: Difference between revisions
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Latest revision as of 16:09, 23 November 2011
Cloning of Cre by subclone 241 to 242
Purpose: to clone 237-pAd5-Cre-GFP
Date: 2011-10-14
Notebook: luyingying
Page:
Clone ID:237
Experimenters: lu yingying
Result
band shows vector
bands show cre
Restart
Date: 2011-10-18 ---
Notebook: luyingying
Page:
Clone ID:237
Experimenters: lu yingying
Result
PCR product
298 plasmid as template
primer:Lenti-Cre-F+Lenti-Cre-R
25 cycles
digest resulte:EcoR1+ Xho1 NEB enzyme 37 for 3 hours
242 plasmid digest as vector
Cloning of Cre by subclone 298 pTriEx-HTNC (Cre) to 242 pAd5-CMV-2-IRES-GFP
Purpose: to clone 237-pAd5-Cre-GFP
Date: 2011-10-25
Notebook: luyingying
Page:
Clone ID:237
Experimenters: lu yingying
Result
band 4,5 show subclone result:EcoR1+ Xho1
band 4 show 242 pAd5-CMV-2-IRES-GFP digest result
band 5 show 298 pTriEx-HTNC (Cre) digest result
band 1-6 show digest detect result:EcoR1+ Xho1
Date: 2011-11-10 ---
Notebook: luyingying
Page:
Clone ID:237
Experimenters: lu yingying
Result
PCR amplify His-Tat-NLS-Cre(298) as temlate, using Cre-EcoR1-F B1 + Cre-BglII-R B2
PCR product with EcoR1 + BglII, digest pAd5-CMV-IRES-GFP with BamH1 + EcoR1
band 1 show insert, band 2 show pAd5-CMV-IRES-GFP(242) vector digest
digest detect:EcoR1+ Not1
Date: 2011-11-16 ---
Notebook: luyingying
Page:
Clone ID:237
Experimenters: lu yingying
Result
PCR amplify His-Tat-NLS-Cre(298) as temlate, using Cre-EcoR1-F2 B1 + Cre-BglII-R B2
PCR product with EcoR1 + BglII, digest pAd5-CMV-IRES-GFP with BamH1 + EcoR1
digest test:EcoR1+Not1