Team:TU-Delft/Notebook/2013/09/19/

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Notebook

June
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30
July
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August
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September
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19th of September


Lab work

1. Restriction digestion is started for pTet cI TT with S+P and with pcI Ulp with X+P. 2. Colony PCR was done for the following:



3. The lysis experiment was carried out on a plate reader for 2 colonies from the pT7 Lysis transformed into BL21 plysS to check whether on inducing the pT7 promoter with IPTG, the cells lyse. The 10 X IPTG stock was made from 1000X IPTG stock (10 µL of 1000X IPTG in 990 µL of LB Broth). The loading of samples on the 96-well plate was done as follows:

Lane Cells (µL) 10 X IPTG (µL) LB Broth (µL)
Lane 1 0 10 90
Lane 2 5 1 94
Lane 3 5 2 93
Lane 4 5 3 92
Lane 5 5 4 91
Lane 6 5 5 90
Lane 7 5 6 89
Lane 8 5 7 88
Lane 9 5 8 87
Lane 10 5 9 86
Lane 11 5 10 85
Lane 12 5 0 95

The results can be seen in the page : Lysis experiment
5. Ligations were done for :
          Lysis Device + pSB1C3
          pTet cI TT + pSB1C3
          pcI Ulp + pSB1C3
6. Reinoculated the colonies from the colony PCR.