Paul.xu at 17:29, 20 April 2012

2012-04-20T17:29:17Z

New page

== '''Cloning of 534-pLVX-EF1-IRES-EGFP''' ==

'''Purpose:''' Cloning of 534-pLVX-EF1-IRES-EGFP by PCR 

'''Date: '''2012-1-13--2012-1-17

'''Notebook:'''

'''Page:'''

'''Clone ID:534'''

'''Experimenters:''' yingying

== '''Results''' ==

[[Image:323 PCR 1-4-2012.JPG]] 

template: pPuroGreen 178

Primer-F EF1-NdeI-F agct GCTAGC c gtgaggctcc ggtgcccg <--- this primer is wrong, it has Nhe1 site, not Nde1 site. should not be used Primer-R EF1-BamH1-R (cttccatttcaggtgtcgtga GGATCC atcg) cgatGGATCCtcacgacacctgaaatggaag 

[[Image:533+534 clone 1-13-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 456px; height: 96px;"
|-
| 
| band 3 insert 
| band 4 vector 
|-
| 
| PCR product 
| 528 pLVX-IRES-EGFP 
|-
| digestion enzymes 
| colspan="2" |            Nde1 + BamH1 
|}

 

=== Subcloning ===

 

 

=== Colony Screening ===

Date:2012-1-17 

Make miniprep and digest with Nde1 + BamH1 

[[Image:533+534 digest detect 1-17-2012.JPG]]

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

* 

== '''Conclusion''' ==

 

=== ===

 

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== '''Cloning of 534-pLVX-EF1-IRES-EGFP''' ==

'''Purpose:''' Cloning of 534-pLVX-EF1-IRES-EGFP by PCR 

'''Date: '''2012-1-18----2012-1-20

'''Notebook:'''

'''Page:'''

'''Clone ID:534'''

'''Experimenters:''' yingying

== '''Results''' ==

[[Image:323 PCR 1-4-2012.JPG]] 

template: pPuroGreen 178

Primer-F EF1-NdeI-F agct GCTAGC c gtgaggctcc ggtgcccg Primer-R EF1-BamH1-R (cttccatttcaggtgtcgtga GGATCC atcg) cgatGGATCCtcacgacacctgaaatggaag 

[[Image:534 clone 1-18-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 456px; height: 96px;"
|-
| 
| band 1 insert 
| band 2 vector 
|-
| 
| PCR product 
| 528 pLVX-IRES-EGFP 
|-
| digestion enzymes 
| colspan="2" |            Nde1 + BamH1 
|}

 

=== Subcloning ===

 

 

=== Colony Screening ===

Date:2012-1-20 

Make miniprep and digest with Nde1 + BamH1 

[[Image:534 digest detect 1-20-2012.JPG]]

 

 

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

* 

== '''Conclusion''' ==

 

=== ===

 

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== '''Cloning of 534-pLVX-EF1-IRES-EGFP''' ==

'''Purpose:''' Cloning of 534-pLVX-EF1-IRES-EGFP by PC

'''Date: '''2012-2-6--2012-2-8

'''Notebook:003732'''

'''Page:'''

'''Clone ID:534'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

Date:2012-2-6

[[Image:534 clone 2012-2-6.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 333px; height: 96px;"
|-
| 
| band 1 
| band2 
|-
| 
| PCR product 
| 528 pLVX-IRES-EGFP 
|-
| digestion enzymes 
| colspan="2" | Nde1 + BamH1 
|}

 

 

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

=== Colony Screening ===

Date: 2012-2-8

Make miniprep and digest with Nde1 + BamH1 

[[Image:534+548 digest detect 2-9-2012.JPG]]

11,13 not right size

 

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

* 

== '''Conclusion''' ==

 

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== '''Cloning of EF1-TA vector''' ==

'''Purpose:''' Cloning of EF1-TA vector

'''Date: '''2012-4-4---2012-4-6

'''Notebook:0'''

'''Page:'''

'''Clone ID:'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

TA clone method 

 

 

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=== Colony Screening ===

Date: 2012-4-6

Make miniprep and digest with Nde1 + BamH1

[[Image:EF1-TA vector digest detect 4-6-2012.JPG]]

 

 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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== '''Cloning of 534-pLVX-EF1-IRES-EGFP ''' ==

'''Purpose:''' Cloning of 534-pLVX-EF1-IRES-EGFP 

'''Date: '''2012-4-17---

'''Notebook:0'''

'''Page:'''

'''Clone ID:534'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

[[Image:534 PCR 4-17-2012.JPG]] 

1101 pFUSE-hIgG1-Fc2-IAB as template

hEF1-HTLV-Nde1-F catgac catatg GGATCTGCGATCGCTCCGGTG

hEF1-HTLV-EcoR1-R (CAAGCTGTGACCGGCGCCTACCTGAGATCA gaattc actaga) tctagtgaattcTGATCTCAGGTAGGCGCCGGTCACAGCTTG [[Image:534 clone 4-18-2012.JPG]] 

 

{| cellspacing="1" cellpadding="1" border="1" style="width: 281px; height: 66px;"
|-
| 
| band 1 
| band 2 
|-
| 
| PCR product 
| 528 pLVX-IRES-EGFP 
|-
| enzymes 
| colspan="2" |          .Nde1+EcoR1 
|}

 

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

=== Colony Screening ===

Date: 2012-4-20

Make miniprep and digest with .Nde1+EcoR1 

[[Image:534_digest_detect_4-20-2012.JPG]]

 

 

 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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[[Category:Cloning]]

534-pLVX-EF1-IRES-EGFP - Revision history

Paul.xu at 17:29, 20 April 2012