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Team:TU-Delft/Notebook/2013/09/19/
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2. Colony PCR was done for the following: <br> | 2. Colony PCR was done for the following: <br> | ||
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| - | 3 | + | 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:<br> |
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| - | The results can be seen in the page : <a href="https://2013.igem.org/Team:TU-Delft/Killswitch#lysis_experiment" >Lysis experiment</a> <br> | + | The results can be seen in the page : <a href="https://2013.igem.org/Team:TU-Delft/Killswitch#lysis_experiment" target="blank";>Lysis experiment</a> <br> |
| - | 5. | + | 5. Ligations were done for : <br> |
| - | | + | Lysis Device + pSB1C3 <br> |
| - | | + | pTet cI TT + pSB1C3 <br> |
| - | | + | pcI Ulp + pSB1C3 <br> |
| - | 6. Reinoculated | + | 6. Reinoculated the colonies from the colony PCR. <br> |
Latest revision as of 09:09, 4 October 2013
Notebook
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| 29 | 30 |
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.
