Paul.xu at 19:10, 12 April 2012

2012-04-12T19:10:56Z

New page

== '''Cloning of 1011-pET30b-FluASeg-7''' ==

'''Purpose:''' Cloning of 1005-pBad24-Tat-sfGFP by PCR

'''Date: '''2012-3-21--2012-

'''Notebook:'''

'''Page:'''

'''Clone ID:1010'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

Date:2012-3-22

[[Image:1011+1017 PCR 3-21-2012.JPG]] 

template:Influenza A virus (A/WSN/1933(H1N1)) segment 7 

FluASeg-7-NdeI-F actagc CAT ATG CAC CAT CAT CAT CAT CAT agtcttctaaccgaggtc

FluASeg-7-Xho1-R (catcggtctcataggcaaatgCTCGAG atcaga) tctgatCTCGAGcatttgcctatgagaccgatg 

 

[[Image:1011+1017+1021clone 3-22-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 312px; height: 96px;"
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| band 1 
| band 2 
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| PCR product 
| pET30b 
|-
| digest enzymes 
| colspan="2" |        Nde1 + Xho1 
|}

 

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=== Colony Screening ===

Date: 2012-3-26 

Make miniprep and digest with Nde1 + Xho1 

[[Image:1011+1017+1021 digest detect 3-26-2012.JPG]]

clone 1,2,3,4 may be right

 

Protein expression in Nico21(DE3)

[[Image:Taq_muts_purification_4_12_2012.jpg]]

[[Image:1011_ET30b-FluASeg-7_expression.jpg]]

 

 

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

* 

== '''Conclusion''' ==

 

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[[Category:Cloning]]

1011-pET30b-FluASeg-7 - Revision history

Paul.xu at 19:10, 12 April 2012