Paul.xu at 19:22, 15 March 2013

2013-03-15T19:22:19Z

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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 

[[Image:PCR gateway cassett 4 17 2012.jpg]] 

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 

[[Image:1062 1063 PCR 5.1.2012.JPG|106x251px|1062 1063 PCR 5.1.2012.JPG]] 

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

[[Image:1062 1063 P V digest 5.1.2012.JPG]] 

Ligate and transform ccdb Survival 

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

[[Image:1062 1063 digest 5.3.2012.JPG]]

 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 

[[Image:1062+393 1063+393 digest 5.8.2012.JPG]] 

 

== '''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 ===

[[Image:1063 PCR 5-14-2012.JPG]] 

The same to 1063 PCR prodedure,

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

[[Image:537+1035+1062+1063clone 5-15-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 407px; height: 83px;"
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| band 5 
| band 4 
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| PCR product 
| 1061 pLVX-CAG-MCS-IRES-EGFP 
|-
| enzymes 
| colspan="2" |            Age1 + Xho1 
|}

 

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=== Colony Screening ===

Date: 2012-5-17

Make miniprep and digest with EcoR1+Xho1 

[[Image:1062+1063 digest detect 5-17-2012.JPG]]

 

c 

== '''Result intepretation/Summary''' ==

*&lt;br&gt;

== '''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

[[Image:1062_1063pcr.JPG‎]] 

digest :

[[Image:Digest_for_1062.JPG‎]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 298px; height: 82px;"
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| band 1 
| band 3,4 
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|   1061pLVX-CAG-MCS-IRES-EGFP 
| PCR 
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| enzymes 
| colspan="2" |        Nco1+ Xho1 
|}

 

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=== Colony Screening ===

Date: 2013

 

 

 

 

== '''Result intepretation/Summary''' ==

* 

== '''Conclusion''' ==

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[[Category:Cloning]]

1062-pLVX-CAG-Dest-IRES-EGFP - Revision history

Paul.xu at 19:22, 15 March 2013