1062-pLVX-CAG-Dest-IRES-EGFP

Cloning of pLVX-CAG-Dest-IRES-EGFP(1062)
Purpose: to clone pLVX-CAG-Dest-IRES-EGFP
Date: 2012-4-19 to 2012-4-
Notebook: Xiaojing Wang
Page: 004228

Clone ID: 1062
Experimenters: Xiaojing Wang

Results

PCR gateway cassette using GW-AgeI-F + GW-Flag-R-Xho1 and pVP16,446 as template

Cut PCR product with Age1 + Xho1
Cut vector 1061 pLVX-CAG-MCS-IRES-EGFP with Age1 + Xho1

ligate and transform ccdbSurvival

5/1/2012

Method 2
PCR using ccdb-attR1-EcoR1-F & ccdb-attR2-Xho1-R and pVP16,446 as template--reducted ccdb gateway

Digest with EcoR1 + Xho1, clone into EcoR1 + Xho1 digested pLVX-CAG-MCS-IRES-EGFP, 1061.

Ligate and transform ccdb Survival

Select 4 clones to mini-prep
1062 plasmids are digested by EcoR1+Xho1

pDonr221_EGFP(393) + 1062-1 make gateway by LR Clonase II.

Transform 393+1062-1 by STBL3, Amp.

Select 4 clones to mini-prep.

1062+393 plasmids are digested by EcoR1+Xho1

Cloning of 1062-pLVX-CAG-Dest-IRES-EGFP

Purpose: Cloning of 1062-pLVX-CAG-Dest-IRES-EGFP

Date: 2012-5-14---

Notebook:0

Page:

Clone ID:1062

Experimenters: yingying

Results

cloning

The same to 1063 PCR prodedure,

PCR gateway cassette using GW-AgeI-F + GW-Flag-R-Xho1 and pVP16,446 as template

	band 5	band 4
	PCR product	1061 pLVX-CAG-MCS-IRES-EGFP
enzymes	Age1 + Xho1

Colony Screening

Date: 2012-5-17

Make miniprep and digest with EcoR1+Xho1

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Result intepretation/Summary

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Conclusion

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Cloning of pLVX-CAG-Dest-IRES-EGFP

Purpose: pLVX-CAG-Dest-IRES-EGFP

Date: 2013-3-

Notebook:

Page:

Clone ID:1062

Experimenters: Shanyi

Results

cloning

Date:2013-3-

PCR gateway cassette using GW-AgeI-F + GW-Flag-R-Xho1 and pVP16,446 as template,

cut PCR product with Age1 + Xho1

Cut vector 1061 pLVX-CAG-MCS-IRES-EGFP with Age1 + Xho1

ligate and transform ccdbSurvival

digest :

	band 1	band 3,4
	1061pLVX-CAG-MCS-IRES-EGFP	PCR
enzymes	Nco1+ Xho1

Colony Screening

Date: 2013

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Conclusion

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