Team:KULeuven/16 September 2008

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Lab Work

Wet Lab

We started the day as we always did - we miniprepped some parts: K145175, I712074+J23078 and C0062+B0015. We also wanted to digest these parts, but we ran out of SpeI. Actually we thought it would be futile to digest them, as the courses start next week and we will not have time to finish new ligations.

We had colonies of the electroporated cells. We did a PCR to test if these cells contained the correct (i.e. ligated) plasmid. K145014+pSB1A2 was definitely alright and K145201, J23109+J23078 and K145253+K145254 seemed OK as well (K145150+B0034+C0062 failed).

We also tried to redo the analysis of the degradation of GFP. At first we didn't detect any fluorescence. So we re-inoculated the cells into fresh LB medium and allowed them to grow for 4 hours. This made sure that the culture was still in de log-phase when we measured fluorescence. After these 4 hours we could detect fluorescence, but we lost our minimal ABT medium and so we couldn't test any further. The final attempt will follow tomorrow!

As we approach the end of our lab work, we also prepared some samples that will be sequenced. This way we can see if our ligations succeeded. The plasmids that are shown to be correct, will then be officially submitted to the registry.

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