Paul.xu at 22:01, 10 October 2012

2012-10-10T22:01:19Z

New page

== '''Cloning of 1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP''' ==

'''Purpose:''' Cloning of 1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP

'''Date: '''2012-8-20---2012-8-22

'''Notebook:0'''

'''Page:'''

'''Clone ID:1272'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

PCR using 207 pLV-TurboGFP as template, and primers TurboGFP-Xho1-F cgatcgctcgagGCCACCATGGAGAGCGACGAGAG TurboGFP-Xba1-R gtctagTCTAGATTATTCTTCACCGGCATCTGCATC 

[[Image:1272+1336PCR 8-20-2012.JPG]]

 

clone into Xho1+ Xba1 digested 1271 pLVX-loxP-TurboRFP-ODC-loxP-TagRFP 

1: PCR product 

2: vector, 1271 pLVX-loxP-TurboRFP-ODC-loxP-TagRFP 

[[Image:1272+1336 clone 8-20-2012.JPG]] 

 

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=== Colony Screening ===

Date: 2012-8-22

Make miniprep and digest with Xho1+ Xba1 

[[Image:1272+1336 digest detect 8-22-2012.JPG]]

clone 1,2,3,4,5,6may be right

 

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

== ==

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

== '''Conclusion''' ==

 

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== '''Cloning of 1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP''' ==

'''Purpose:''' Cloning of 1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP

'''Date: '''2012-9-21---2012-9-24

'''Notebook:0'''

'''Page:'''

'''Clone ID:1272'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

PCR using 207 pLV-TurboGFP as template, and primers TurboGFP-Xho1-F cgatcgctcgagGCCACCATGGAGAGCGACGAGAG TurboGFP-Xba1-R gtctagTCTAGATTATTCTTCACCGGCATCTGCATC 

[[Image:1272 PCR 9-21-2012.JPG]] 

 

clone into Xho1+ Xba1 digested 1271 pLVX-loxP-TurboRFP-ODC-loxP-TagRFP 

1: PCR product 

2: vector, 1271 pLVX-loxP-TurboRFP-ODC-loxP-TagRFP 

[[Image:1348+1272 clone 9-21-2012.JPG]] 

 

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=== Colony Screening ===

Date: 2012-9-23

Make miniprep and digest with Xho1+ Xba1 

no insert

 

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

== ==

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

== '''Conclusion''' ==

 

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= = '''Cloning of '''1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP =

'''Purpose:''' Cloning of 1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP

'''Date: '''2012-9-27---

'''Notebook:0'''

'''Page:'''

'''Clone ID:1272'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

[[Image:1272_clone_9-27-2012.JPG]] 

 

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| band 1 
| band 2 
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| 1402,pLVX-TurboGFP 
| 1271,pLVX-loxP-TurboRFP-ODC-loxP-TagRFP 
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| enzymes 
| colspan="2" |             Xho1+ Xba1 
|}

 

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=== Colony Screening ===

Date: 2012/9/29

 

digest plasmid with Xho1+ Xba1

 

[[Image:1376 1272 restrition digestion 9 29 2012.jpg]] 

 

 

10/7/2012

Digest with Xba1+Xho1

[[Image:1272 2012 10 7.jpg]]

Transfection 10/9/2012, 10 ul miniprep DNA 

photo on 10/10/2012, 24 hrs after transfection

grow clone # 2 for midiprep-->stable cell line

[[Image:1272 transfection 10 10 2012.jpg]]

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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[[Category:Cloning]]

1272-pLVX-loxP-TurboRFP-ODC-loxP-TurboGFP - Revision history

Paul.xu at 22:01, 10 October 2012