Template:Team:Groningen/CONTENT/LOGBOOK/MSgg
MSgg
In this experiment several components of MSgg medium were tested for being the cause of the cloudiness of the solution.
Find out what causes the cloudiness of the MSgg medium.
To obtain a clear MSgg solution, MOPS must be used instead of MOPS sodium salt.
<a class="postscriptum protocol" href="https://2015.igem.org/Team:Groningen/Protocols_and_Protocols/No_protocol">None</a>
15:45, 21 July 2015 - 16:15, 21 July 2015
In order to determine whether the MSgg was cloudy because of bacterial growth, first the OD at 600 nm was measured, and found to be 0.58. Of the MSgg 2 mL was put in the shaking incubator at 37 °C and 2 mL was put in the fridge at 4 °C to see if the OD would increase.
11:50, 22 July 2015 - 12:25, 22 July 2015
The OD at 600 nm for both the 4 °C and 37 °C sample was measured, and a difference of only 0.005 was found. After 20 hours of growth, this is negligible, so the cloudiness was not caused by bacterial growth. New MSgg was made, but again it was a cloudy solution. It was found out that too little buffer was added, 0.7 mL instead of 7 mL, so this could explain why the medium was wrong. The medium was used in the drip flow reactor to see what would happen anyway.
13:30, 24 July 2015 - 16:00, 24 July 2015
MOPS sodium salt was tested by making a solution of MSgg in a 100 mL flask with adding 1 mL 100x KxHyPO4 and 1 mL 100x MgCl.2H2O, added up to 88 mL with demiwater instead of 100 mL to see if that was the problem. However, after autoclaving, already the solution was cloudy even without MOPS added. 200 mL new MSgg was made, this time using MOPS instead of MOPS sodium salt. This appeared to have been the problem, as this time the solution did not become cloudy anymore.