Usually it is recommended to add glucose after sterilization of media.
Can anyone tell me, then how should we add glucose in the media?
We usually work on litres of media (4 to 8 L), for which we need 30 to 50 g glucose.
If we add solid glucose then there are chances of contamination.
Should we use microfilters?
Microfilters are very expensive.
Then what others?
In our experiments, we do not care how much glucose is oxidizing on heating in autoclave because our concern is to obtain enough growth of the culture for biotransformation.
You can make 40% glucose and sterilize it by filtering it through 0.2micron filter. If you make a liter of glucose solution, one filter (designed to fit on bottles) will do well. This would be enough for 10 batches of your medium, reducing cost of filters/batch.
Alternatively, you can autoclave the stock glucose solution on liquid cycle but avoid letting it sit too long in the autoclave after the cycle is complete. For many cells, autoclaved glucose works fine.
Generally you can do it by preparing higher concentration glucose stock and get it sterilized by filter sterilisation.
Filter sterilization is the option. However, sterilization (15 lb/ 15 minutes) is not affecting the properties and could be carried out, if you don't prolong the process.
You can either filter sterilise or autoclave a concentrated glucose solution. If you choose the latter, be careful: concentrated solutions of glucose have a quite higher viscosity compared to common aqueous solutions and you will need a larger head space in your bottle to avoid spilling it.
Hello, you mention the concentration of glucose is 30-50 g per liter or the total culture medium prepare. You can dissolve the glucose in 150 ml sterile water and filter through a millipore membrane, and that is the medium and add to a sterile laminar flow hood or directly the bioreactor with a peristaltic pump . The second way is : 150 or 200 ml autoclave sterilization at 15 lb / cm2 for 15 min and adicionas to the culture medium . The fact of putting glucose together with the other components of the medium, is primarily nitrogen source as it reacts with compounds forming glucasa inihiben microbial growth ( Maillard reactions ) if your medium does not have high concentrations of amino acids , proteins , ammonium salts put no problem glucose culture medium and esterilices together .
Yes, filter sterilisation is quite helpful, we have been using syringe filter of 0.22 micrometer size.
- Badges
- Science topic