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Team:DTU-Denmark/Notebook/4 July 2013
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(→Cultivation of TOP10 cells in SOB) |
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| - | =301= | + | {{:Team:DTU-Denmark/Templates/StartPage|4 July 2013}} |
| - | == | + | Navigate to the [[Team:DTU-Denmark/Notebook/3_July_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/5_July_2013|Next]] Entry |
| + | =Lab 301= | ||
| + | <hr/> | ||
| + | ==Main purpose== | ||
| + | <hr/> | ||
We want to view our cells throught a fluorescent microscope, in order to see if they have expressed GFP and RPF. | We want to view our cells throught a fluorescent microscope, in order to see if they have expressed GFP and RPF. | ||
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Setup an overnight culture of the 2 successful transformants so we can both MidiPrep and isolate periplasmic proteins tomorrow. | Setup an overnight culture of the 2 successful transformants so we can both MidiPrep and isolate periplasmic proteins tomorrow. | ||
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==Results== | ==Results== | ||
| + | <hr/> | ||
We managed to see expressed GFP and RFP | We managed to see expressed GFP and RFP | ||
| + | =Lab 115= | ||
| + | <hr/> | ||
| + | ==Main purpose== | ||
| - | + | Preparation for making competent cells based on the [http://parts.igem.org/Help:Protocols/Competent_Cells protocol]. | |
| - | + | ||
| - | + | ||
| - | Preparation for making competent cells based on the | + | |
==Who was in the lab== | ==Who was in the lab== | ||
<hr/> | <hr/> | ||
| - | Henrike, Ariadni, | + | Henrike, Ariadni, Kasia, Natalia |
==Procedure== | ==Procedure== | ||
| + | <hr/> | ||
| - | + | ===Preparation of SOB medium=== | |
| - | ==Preparation of SOB medium== | + | |
| - | + | ||
Total volume 1 L | Total volume 1 L | ||
* 5 g yeast extract | * 5 g yeast extract | ||
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* 0.58 g NaCl | * 0.58 g NaCl | ||
* 0.19 g KCl | * 0.19 g KCl | ||
| - | * 2.41 g | + | * 2.41 g MgSO<sub>4</sub> |
| - | ==Preparation of CCMB80 buffer (500 ml)== | + | ===Preparation of CCMB80 buffer (500 ml)=== |
* KOAc :5 ml (0.98 gr) | * KOAc :5 ml (0.98 gr) | ||
| - | * CaCl2. | + | * CaCl2.2H<sub>2</sub>O (5.9 g) |
| - | * MnCl2. | + | * MnCl2.4H<sub>2</sub>O (2 g) |
| - | * MgCl2. | + | * MgCl2.6H<sub>2</sub>O (1 g) |
* 10% glycerol (50 ml) | * 10% glycerol (50 ml) | ||
Adjustment of pH (6.37) with 0.1 HCl | Adjustment of pH (6.37) with 0.1 HCl | ||
| - | ==Preparation of LB agar== | + | ===Preparation of LB agar=== |
== Conclusion from today== | == Conclusion from today== | ||
| - | + | <hr/> | |
Cultivation of TOP10 cells in SOB | Cultivation of TOP10 cells in SOB | ||
* 1 loop of TOP10 cells (from 14.06.2013) | * 1 loop of TOP10 cells (from 14.06.2013) | ||
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Incubate for 16 hours in 24 degrees and 37 degrees. | Incubate for 16 hours in 24 degrees and 37 degrees. | ||
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| + | Navigate to the [[Team:DTU-Denmark/Notebook/3_July_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/5_July_2013|Next]] Entry | ||
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| + | {{:Team:DTU-Denmark/Templates/EndPage}} | ||
Latest revision as of 11:18, 4 October 2013
4 July 2013
Contents |
Lab 301
Main purpose
We want to view our cells throught a fluorescent microscope, in order to see if they have expressed GFP and RPF.
Hopefully we will see GFP in the periplasm and RFP inside the cell.
Setup an overnight culture of the 2 successful transformants so we can both MidiPrep and isolate periplasmic proteins tomorrow.
Results
We managed to see expressed GFP and RFP
Lab 115
Main purpose
Preparation for making competent cells based on the [http://parts.igem.org/Help:Protocols/Competent_Cells protocol].
Who was in the lab
Henrike, Ariadni, Kasia, Natalia
Procedure
Preparation of SOB medium
Total volume 1 L
- 5 g yeast extract
- 20.05 g tryptone
- 0.58 g NaCl
- 0.19 g KCl
- 2.41 g MgSO4
Preparation of CCMB80 buffer (500 ml)
- KOAc :5 ml (0.98 gr)
- CaCl2.2H2O (5.9 g)
- MnCl2.4H2O (2 g)
- MgCl2.6H2O (1 g)
- 10% glycerol (50 ml)
Adjustment of pH (6.37) with 0.1 HCl
Preparation of LB agar
Conclusion from today
Cultivation of TOP10 cells in SOB
- 1 loop of TOP10 cells (from 14.06.2013)
- 10 ml of SOB medium
Incubate for 16 hours in 24 degrees and 37 degrees.
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