1063-pLVX-CAG-Dest-IRES-mCherry

Cloning of pLVX-CAG-Dest-IRES-mCherry(1063)
Purpose: to clone pLVX-CAG-Dest-IRES-mCherry
Date: 2012-4-12 to 2012-4-
Notebook: Xiaojing Wang
Page: 004224,004215

Clone ID: 1063
Experimenters: Xiaojing Wang

Results

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

4/17/2012:

Cut PCR product with Age1 + Xho1
Cut vector 1060 pLVX-CAG-MCS-CMV-mCherry with Age1 + Xho1

ligate and transform ccdbSurvival

Select 1 clone to do PCR screening 4/19/2012

mini-prep, digest 4/20/2012

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-mCherry, 1060.

Ligate and transform ccdb Survival

Select 4 clones to mini-prep
1063 plasmids are digested by EcoR1+Xho1

pDonr221_EGFP(393) + 1063-1 make gateway by LR Clonase II.

Transform 393+1063-1 by STBL3, Amp.

Select 1 clones to mini-prep.

1063+393 plasmid is digested by EcoR1+Xho1

Cloning of 1063-pLVX-CAG-Dest-IRES-mCherry

Purpose: Cloning of 1063-pLVX-CAG-Dest-IRES-mCherry

Date: 2012-5-14---

Notebook:0

Page:

Clone ID:1063

Experimenters: yingying

Results

cloning

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

	band 5	band 6
	PCR product	1060,pLVX-CAG-MCS-CMV-mCherry
enzymes	Age1 + Xho1

Colony Screening

Date: 2012-5-17

Make miniprep and digest with EcoR1+Xho1

c

Result intepretation/Summary

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Conclusion

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