534-pLVX-EF1-IRES-EGFP

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Cloning of 534-pLVX-EF1-IRES-EGFP

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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

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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



 band 3 insert
 band 4 vector

 PCR product
 528 pLVX-IRES-EGFP
digestion enzymes
            Nde1 + BamH1


Subcloning

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Colony Screening

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Date:2012-1-17

Make miniprep and digest with Nde1 + BamH1

Result intepretation/Summary

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Conclusion

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[edit]




Cloning of 534-pLVX-EF1-IRES-EGFP

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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

[edit]


template: pPuroGreen 178

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



 band 1 insert
 band 2 vector

 PCR product
 528 pLVX-IRES-EGFP
digestion enzymes
            Nde1 + BamH1


Subcloning

[edit]



Colony Screening

[edit]

Date:2012-1-20

Make miniprep and digest with Nde1 + BamH1



Result intepretation/Summary

[edit]

Conclusion

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[edit]



Cloning of 534-pLVX-EF1-IRES-EGFP

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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

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cloning

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Date:2012-2-6



band 1
 band2

 PCR product
 528 pLVX-IRES-EGFP
digestion enzymes
 Nde1 + BamH1




Colony Screening

[edit]

Date: 2012-2-8

Make miniprep and digest with Nde1 + BamH1

11,13 not right size



Result intepretation/Summary

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  • <br>

Conclusion

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<br>


<br>




Cloning of EF1-TA vector

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Purpose: Cloning of EF1-TA vector

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

Notebook:0

Page:

Clone ID:

Experimenters: yingying

Results

[edit]

cloning

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TA clone method




Colony Screening

[edit]

Date: 2012-4-6

Make miniprep and digest with Nde1 + BamH1



Result intepretation/Summary

[edit]
  • &lt;br&gt;

Conclusion

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&lt;br&gt;


&lt;br&gt;


<br>




Cloning of 534-pLVX-EF1-IRES-EGFP

[edit]

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

Date: 2012-4-17---

Notebook:0

Page:

Clone ID:534

Experimenters: yingying

Results

[edit]

cloning

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1101 pFUSE-hIgG1-Fc2-IAB as template

hEF1-HTLV-Nde1-F
catgac catatg GGATCTGCGATCGCTCCGGTG

hEF1-HTLV-EcoR1-R
(CAAGCTGTGACCGGCGCCTACCTGAGATCA gaattc actaga)
tctagtgaattcTGATCTCAGGTAGGCGCCGGTCACAGCTTG



 band 1
 band 2

 PCR product
528 pLVX-IRES-EGFP
enzymes
          .Nde1+EcoR1



Colony Screening

[edit]

Date: 2012-4-20

Make miniprep and digest with .Nde1+EcoR1




Result intepretation/Summary

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  • &lt;br&gt;

Conclusion

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&lt;br&gt;


&lt;br&gt;


<br>




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