237-pAd5-Cre-GFP

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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