Difference between revisions of "Part:BBa K1111012:Experience"

Revision as of 22:15, 2 October 2013

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_K1111012

This part can be used in any experience with biotin. Biotinylation of biological material is widely used and quick to do. Once biotinylated, the molecule will be able to attach to the streptavidin expressed in bacteria. For us, the material was nanoparticles, that we nicely biotinylated ([http://2013.igem.org/Team:EPF_Lausanne/Nanoparticles nanoparticles protocol and usage])

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 are shown here : File:Team-EPF-Lausanne ClustalW2 VR col1&2 ISA.pdf

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

UNIQc069a3653372253d-partinfo-00000000-QINU UNIQc069a3653372253d-partinfo-00000001-QINU

@@ Line 8: / Line 8: @@
 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 are shown here :
-[[Image: CADSequ.jpg|thumb|600px|center|Figure 2: Sequencing result of the Cad promoter using one foreward Primer that binds directly to the beginning of the promoter ]]
+[[File: Team-EPF-Lausanne_ClustalW2_VR_col1%262_ISA.pdf|thumb|600px|center|Figure 2: Sequencing result of the Cad promoter using one foreward Primer that binds directly to the beginning of the promoter ]]
 ==Microscopy==