Difference between revisions of "Part:BBa K1456001:Design"
 
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<center>[[File:ATOMS-tpa_results1.png|400px|]]</center><br/>
 
<center>[[File:ATOMS-tpa_results1.png|400px|]]</center><br/>
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== '''Cloning''' ==
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===Cloning===
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[[File:https://static.igem.org/mediawiki/2014/f/f4/ATOMS-tpa_results3.png]]
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<center>[[File:ATOMS-tpa_results3.png|400px|]]</center><br/>
 
*Through the primers that we ordered and the cDNA, we acquired tPA inserts. The insert was, as expected, portraying that the base length was around 1700 bp.
 
*Through the primers that we ordered and the cDNA, we acquired tPA inserts. The insert was, as expected, portraying that the base length was around 1700 bp.
  
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<partinfo>BBa_K1456001 SequenceAndFeatures</partinfo>
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<center>[[File:ATOMS-tpa_results5.png|400px|]]</center><br/>
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*The specified primers were put into colony PCR. As it can be seen in the results above, a right insert was not achieved. This process was repeated several times but no result was achieved. Seeing that ligation did not provide a solution to the problem, synthetically produced inserts were ordered.
  
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<center>[[File:ATOMS-tpa_results7.png|400px|]]</center><br/>
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*Colony PCR was applied to the inserts acquired from transformation and ligation. As it can be seen in the figure above, the second colony contains the appropriate base length in respect to the ladder.
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<partinfo>BBa_K1456001 SequenceAndFeatures</partinfo>
  
 
===Design Notes===
 
===Design Notes===
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None
 
  
 
===Source===
 
===Source===

Latest revision as of 14:28, 21 October 2014

Human Tissue Plasminogen Activator (htPA)

ATOMS-tpa results1.png

Cloning

ATOMS-tpa results3.png

  • Through the primers that we ordered and the cDNA, we acquired tPA inserts. The insert was, as expected, portraying that the base length was around 1700 bp.


ATOMS-tpa results5.png

  • The specified primers were put into colony PCR. As it can be seen in the results above, a right insert was not achieved. This process was repeated several times but no result was achieved. Seeing that ligation did not provide a solution to the problem, synthetically produced inserts were ordered.


ATOMS-tpa results7.png

  • Colony PCR was applied to the inserts acquired from transformation and ligation. As it can be seen in the figure above, the second colony contains the appropriate base length in respect to the ladder.



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 5
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 119
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 958

Design Notes

Source

We clonned this part from human hepatocellular carcinoma cell genome cDNA.

References