Difference between revisions of "Part:BBa K175001:Experience"

 
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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===Applications of BBa_K175001===
 
===Applications of BBa_K175001===
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===Characterization of BBa_K175001===
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A conjugation test was done using the TUDelft iGEM 2009 [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation protocol] to verify that expression of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K175001 trbK] blocked conjugation.
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'''Donors''': [http://www.cbs.knaw.nl/databases/nccb/search_bac_plas.aspx NCCBNr# 2350] cells with R751 (TRI)<br>
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'''Receivers''': DH5 &alpha; cells expressing [https://parts.igem.org/wiki/index.php?title=Part:BBa_K175001 trbK] (CAM) <br>
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'''Transconjugants''': DH5 &alpha; cells with R751 and expressing [https://parts.igem.org/wiki/index.php?title=Part:BBa_K175001 trbK] (TRI+CAM)<br><br>
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'''Receivers Control''': DH5 &alpha; cells with [https://parts.igem.org/Part:BBa_I13522 I13522] (AMP) <br>
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'''Transconjugants Control''': DH5 &alpha; cells with R751 and [https://parts.igem.org/Part:BBa_I13522 I13522] (TRI+AMP)<br>
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The positive control test used the same donors but different receivers, as detailed above.
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The following table shows the calculated conjugation efficiencies (raw data [http://2009.igem.org/Team:TUDelft/19_August_2009#Calin here]):
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{| border="1" align="center"
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|- align="center"
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| Set # || Conjugation Efficinecy
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|- align="center"
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| trbK 1 || 4.49E-6
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|- align="center"
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| trbK 2 || 2.89E-6
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|- align="center"
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| control || 0.031
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|- align="center"
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|}
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[[Image:TUD_aug18_trbK_efficinecy.png|center]]
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As the graph above shows, expression of trbK reduces the conjugation efficiency by four orders of magnitude. This can be used as receiver side control over conjugation.
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===User Reviews===
 
===User Reviews===

Latest revision as of 21:19, 19 October 2009

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K175001

Characterization of BBa_K175001

A conjugation test was done using the TUDelft iGEM 2009 [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation protocol] to verify that expression of trbK blocked conjugation.

Donors: [http://www.cbs.knaw.nl/databases/nccb/search_bac_plas.aspx NCCBNr# 2350] cells with R751 (TRI)
Receivers: DH5 α cells expressing trbK (CAM)
Transconjugants: DH5 α cells with R751 and expressing trbK (TRI+CAM)

Receivers Control: DH5 α cells with I13522 (AMP)
Transconjugants Control: DH5 α cells with R751 and I13522 (TRI+AMP)

The positive control test used the same donors but different receivers, as detailed above.

The following table shows the calculated conjugation efficiencies (raw data [http://2009.igem.org/Team:TUDelft/19_August_2009#Calin here]):

Set # Conjugation Efficinecy
trbK 1 4.49E-6
trbK 2 2.89E-6
control 0.031


TUD aug18 trbK efficinecy.png


As the graph above shows, expression of trbK reduces the conjugation efficiency by four orders of magnitude. This can be used as receiver side control over conjugation.


User Reviews

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