Part:BBa_K1111013:Experience

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_K1111013

As said in the main page, this part is a negative control with a lower Kd compared to BBa_K1111012 and BBa_K1111014.

Sequencing

We sequenced this part once the Gibson assembly made. To do so, we used primers for iGEM sites VF2 and VR, in order to sequence all that was inserted in the backbone pSB1C3. The sequencing results were aligned to the theoretical coding sequences using ClustalW2 program [http://www.ebi.ac.uk/Tools/msa/clustalw2/]. In the following pdf, the starts mean 100% matching sequences.
VF2 sequencing results of INP_Streptavidin Dead:
File:Team-EPF-Lausanne ClustalW2 VF2 ISD.pdf
VR sequencing results of INP_Streptavidin Dead:
File:Team-EPF-Lausanne ClustalW2 VR ISD.pdf
Note: you can notice that there is no overlap between the two sequencing results but thanks to another forward primer (5'- AATAATATGGCCGACCATTG -3') we designed, we were able to confirm the sequences.

Microscopy

Also, we transformed cells and used them to do microscopy. Two experiments were done, one with a fluorescent antibody against streptavidin and the other one with a fluorescent biotin. We did both in case the antibody would be too big, since biotin is a small molecule best suit to cross glycocalix.

User Reviews

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