Coding

Part:BBa_K2719002:Experience

Designed by: Karla Soto Blas   Group: iGEM18_TecCEM   (2018-08-08)


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Applications of BBa_K2719002

To confirm the presence of Tenascin, it was cloned in pSB1C3 using EcoRI and PstI for the restriction and T4 ligase for the ligation. After that, it was transformed in Escherichia coli DH5a. (Figure 2)

T--TecCEM--TCD5Colonies.png

Figure 1. Transformed Tenascin 5 Domain V colonies

To prove the presence of the plasmid in E. coli it was necessary to create an agarose gel. Because polyproline linker wasn’t binded in this fusion protein, the bands presented in the gel were less heavy. (Figure 2).

After that, another agarose gel was made for the restriction parts to confirm the presence of the insert (figure 3).

T--TecCEM--FusGCT.png

Figure 2.GST and Tenascin V Fusion agarose gel at 0.85% with GelRed: MW) 1Kb plus from NEB; 3) GST and Tenascin V Fusion (BBa_K2719002) V'; 4) GST and Tenascin V Fusion (BBa_K2719002) V

T--TecCEM--ResK2719002.png

Figure 3.GST and Tenascin V Fusion restriction with NotI agarose gel at 0.85% with GelRed: MW) 1Kb plus from NEB; 5) GST and Tenascin V Fusion (BBa_K2719002) V'; 7) GST and Tenascin V Fusion (BBa_K2719002) V

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