Help: how to increase carbon dioxide level ? - (Mar/01/2011 )
Hi all
I'm new at this forum.
For the moment I'm working on Sodalis glossinidius. I'm trying to insert a foreign DNA construct via the lambda-Red recombination system. Unfortunately without any success so far: no colonies on agar plates after transformation. Sodalis grows very slowly under microaerophilic (5% O2; 95% CO2)conditions. I've found out that de CampyGen atmospehere generation system (Oxoid) I use only generates 9 - 13% CO2; far less than the optimal level (95%). This may be the reason why the colonies won't grow. Does anybody know how I can increase the CO2 level?
Many thanks.
Have you tried a candle jar? Depending on whether it is the microaerophilic or capnophilic characteristics of the organism that control growth this may work.
You could get a CO2 cylinder and an O2 cylinder and a couple of regulators ...
The gas cylinder is the best option.
But if that is not available, how about adding dry ice?
Thanks all for the commments!
The problem is that the lab I work in doesn't have CO2/O2 cylinders.
How should the dry ice work?
Timasi on Wed Mar 2 08:57:42 2011 said:
Thanks all for the commments!
The problem is that the lab I work in doesn't have CO2/O2 cylinders.
How should the dry ice work?
Dry ice = carbon dioxide... its just the solid form.
I think this is the best option for you since you dont have the utilities to use a CO2 cilinder.
I assume that you've seen the following paper:
Appl Environ Microbiol. 2011 Mar;77(5):1918-20. Epub 2011 Jan 7.
Lambda Red-Mediated Genetic Modification of the Insect Endosymbiont Sodalis glossinidius.
Pontes MH, Dale C.
Did you contact the authors and ask for advice?