Team:TU-Delft/Notebook/2013/09/19/
From 2013.igem.org
Notebook
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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.