Difference between revisions of "Part:BBa K1709002:Experience"
(→‎User Reviews)
Line 38: Line 38:
 
|};
 
|};
 
<!-- End of the user review template -->
 
<!-- End of the user review template -->
−
<p>To characterize the CheZ-GFP BioBrick, the fragment containing a RBS was cloned directly after a strong promotor (BBa_J23101). The first figure shows the gel right before ligation.<br/>
 
−
The colonies were checked by restriction mapping using BcuI and PstI (results not shown). The DNA sequence was also confirmed by DNA sequencing. Results can be provided by email. The presence of colonies expressing GFP proves that the plasmid was designed and cloned correctly.<br/>
 
−
<br/>
 
−
Further DNA sequencing proved the correct sequence, as designed. </p>
 
  
−
 
−
[[File:KU_Leuven_GelPurification.jpeg|500px|center|]]
 
−
 
−
<p>Gel after purification. Lanes 2-5: insert (1400bp). Lane 6: linearized vector. Lanes 7-10 : insert. Lane 11: linearized vector </p>
 
−
<br/>
 
−
 
−
[[File:KU_Leuven_fluorescence.jpg|500px|center|]]
 
−
 
−
<p>GFP is expressed in the cells. This confirms the correct construction of the BioBrick </p>
 
−
<br/>
 
−
 
−
<p>Further characterization could be done by transforming the <i>cheZ</i> knockout Keio strain with this plasmid. These cells should then regain their possibility to swim.</p>
 
−
<br/>
 
−
<br/>
 
 
<!-- DON'T DELETE --><partinfo>BBa_K1709002 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K1709002 EndReviews</partinfo>

Revision as of 14:52, 26 September 2015

The sequence was verified by restriction mapping and PCR and the presence of colonies expressing GFP proves that the plasmid was designed and cloned correctly.

Further DNA sequencing proved the correct sequence, as designed.

To characterize the CheZ-GFP BioBrick, the fragment containing a RBS was cloned directly after a strong promotor (BBa_J23101). The first figure shows the gel right before ligation.
The colonies were checked by restriction mapping using BcuI and PstI (results not shown). The DNA sequence was also confirmed by DNA sequencing. Results can be provided by email. The presence of colonies expressing GFP proves that the plasmid was designed and cloned correctly.

Further DNA sequencing proved the correct sequence, as designed.


KU Leuven GelPurification.jpeg

Gel after purification. Lanes 2-5: insert (1400bp). Lane 6: linearized vector. Lanes 7-10 : insert. Lane 11: linearized vector


KU Leuven fluorescence.jpg

GFP is expressed in the cells. This confirms the correct construction of the BioBrick


Further characterization could be done by transforming the cheZ knockout Keio strain with this plasmid. These cells should then regain their possibility to swim.



Applications of BBa_K1709002

User Reviews

UNIQc7e828000123b993-partinfo-00000000-QINU

UNIQc7e828000123b993-partinfo-00000001-QINU