Paul.xu at 18:08, 4 December 2012

2012-12-04T18:08:54Z

New page

== '''Cloning of 1426-pCMV5-TPA-Dest2-(3c)-Fc-WPRE-OriP''' ==

'''Purpose:''' Cloning of 1426-pCMV5-TPA-Dest2-(3c)-Fc-WPRE-OriP

'''Date: '''2012-10-23---2012-10-25

'''Notebook:0'''

'''Page:'''

'''Clone ID:1426'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

[[Image:1425+1426 PCR 10-23-2012.JPG]] 

PCR Gateway Cassette out from the new cassette  1336 pCMV5-Fc(3C)-Dest-WPRE 

GW-Nhe1-F2 +GW-Not1-Kpn1-R 

 

[[Image:1425+1426 clone 10-23-2012.JPG]] 

{| width="397" cellspacing="1" cellpadding="1" border="1"
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| lane3 
| lane4 
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| PCR product 
| 1391,pCMV5-TPA-TagGFP-(3c)-Fc-WPRE-OriP 
|-
| enzymes 
| colspan="2" |    Nhe1 + Kpn1 
|}

 

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|}

=== Colony Screening ===

Date: 2012-10-25

Make miniprep and digest with Nhe1 + Kpn1 

[[Image:1425+1426+1433 digest detect 10-25-2012.JPG]]

clone 1,2,3,4,6 need to sequence

 

'''Result intepretation/Summary'''

== ==

*&amp;lt;br&amp;gt;

== '''Conclusion''' ==

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== '''Cloning of 1426-pCMV5-TPA-Dest2-(3c)-Fc-WPRE-OriP''' ==

'''Purpose:''' Cloning of 1426-pCMV5-TPA-Dest2-(3c)-Fc-WPRE-OriP

'''Date: '''2012-11-26---2012-12-4

'''Notebook:0'''

'''Page:'''

'''Clone ID:1426'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

PCR Gateway Cassette out from the new cassette  1336 pCMV5-Fc(3C)-Dest-WPRE 

GW-Nhe1-F2 +GW-Not1-Kpn1-R 

[[Image:1426_PCR_11-26-2012.JPG]]

 

[[Image:1424+1426+1472+1473+1474+1475+1476 clone 11-26-2012.JPG]] 

{| width="397" cellspacing="1" cellpadding="1" border="1"
|-
| 
| lane3 
| lane4 
|-
| 
| PCR product 
| 1391,pCMV5-TPA-TagGFP-(3c)-Fc-WPRE-OriP 
|-
| enzymes 
| colspan="2" |    Nhe1 + Kpn1 
|}

 

{| cellspacing="1" cellpadding="1" border="1"
|-
| 
|}

=== Colony Screening ===

Date: 2012-11-30

PCR Screening: CMV5+GW-Not1-Kpn1-R

[[Image:1426 PCR Screen 11-30-2012.JPG]] 

Grow 2,5,8,11 

2012-12-3 mini prep and digest with Nhe1 + Kpn1 

[[Image:1426 digest detect 12-4-2012.JPG]]

1,2,3,4 the same with (2,5,8,11 above),

need to sequencing

 

 

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

*&amp;lt;br&amp;gt;

== '''Conclusion''' ==

&amp;lt;br&amp;gt;

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&amp;lt;br&amp;gt;

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[[Category:Cloning]]

1426-pCMV5-TPA-Dest2-(3c)-Fc-WPRE-OriP - Revision history

Paul.xu at 18:08, 4 December 2012