Difference between revisions of "Team:UGent Belgium/LabNotebook"
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| − | We made a strong expression vector (pXS) by restriction of the linearized plasmid backbone pSB1C3 and insertion of gblock made up of promoter-RBS-cloning site-terminator-terminator (cut with EcoRI and PstI) | + | We made a strong expression vector (pXS) by restriction of the linearized plasmid backbone pSB1C3 and insertion of gblock made up of promoter-RBS-cloning site-terminator-terminator (cut with EcoRI and PstI). |
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| − | Our first attempts to isolate the inaZ gene from <i>P. syringae</i> and the inaX gene from <i>X. campestris</i>failed | + | Our first attempts to isolate the inaZ gene from <i>P. syringae</i> and the inaX gene from <i>X. campestris</i> failed. |
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| − | We transformed the strong expression vector (cf. 29/08) by electroshock into <i>E. coli</i> TOP10 | + | We transformed the strong expression vector (cf. 29/08) by electroshock into <i>E. coli</i> TOP10. |
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| − | Second attempt to isolate the inaZ gene from <i>P. syringae</i> and the inaX gene from <i>X. campestris</i>. InaX failed, but inaZ was successful | + | Second attempt to isolate the inaZ gene from <i>P. syringae</i> and the inaX gene from <i>X. campestris</i>. InaX failed, but inaZ was successful. |
</li> | </li> | ||
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| − | Transformation of the strong expression vector by restriction and ligation was not very successful, we only had a few colonies. The backbone amplification using CPEC was successful (cf 30/08). | + | Transformation of the strong expression vector by restriction and ligation was not very successful, we only had a few colonies. The backbone amplification using CPEC was successful (cf. 30/08). |
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Revision as of 20:11, 6 October 2016
Lab Notebook
- August 29:
- We made a strong expression vector (pXS) by restriction of the linearized plasmid backbone pSB1C3 and insertion of gblock made up of promoter-RBS-cloning site-terminator-terminator (cut with EcoRI and PstI).
- Our first attempts to isolate the inaZ gene from P. syringae and the inaX gene from X. campestris failed.
- August 30:
- We transformed the strong expression vector (cf. 29/08) by electroshock into E. coli TOP10.
- As a backup strategy we also used CPEC with the backbone from the K584027 plasmid to generate a strong expression vector.
- August 31:
- Second attempt to isolate the inaZ gene from P. syringae and the inaX gene from X. campestris. InaX failed, but inaZ was successful.
- Transformation of the strong expression vector by restriction and ligation was not very successful, we only had a few colonies. The backbone amplification using CPEC was successful (cf. 30/08).
