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Team:DTU-Denmark/Notebook/4 July 2013
From 2013.igem.org
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==Results== | ==Results== | ||
We managed to see expressed GFP and RFP | We managed to see expressed GFP and RFP | ||
| + | |||
| + | |||
| + | =115= | ||
| + | ==main purpose== | ||
| + | |||
| + | Preparation for making competent cells based on the protocol ([http://parts.igem.org/Help:Protocols/Competent_Cells]). | ||
| + | |||
| + | ==Who was in the lab== | ||
| + | <hr/> | ||
| + | Henrike, Ariadni, Kashia, 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. | ||
Revision as of 20:16, 7 July 2013
Contents |
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.
Procedure
Results
We managed to see expressed GFP and RFP
115
main purpose
Preparation for making competent cells based on the protocol ([http://parts.igem.org/Help:Protocols/Competent_Cells]).
Who was in the lab
Henrike, Ariadni, Kashia, 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.
