1. I am trying to culture methanotrophs using the mineral medium (DSMZ 1007 Mineral Medium) . The instruction says to grow it under gas phase of 20% methane in air. By culturing this using bottle and butyl stopper (i dont have anaerobic jar), how do I calculate this 20% methane in air?
2. The instruction also said some methanotroph could be grown on either 0.5-1.0% methanol. Can someone explain this to me too? Does it mean, I add 0.5-1.0% into the mineral medium?
Hi,
The best possibility is to order a ready made gas mixture (20% methane in air) and use is as gas phase. The consumption of methane can be followed by pressure and gas chromatography analysis of the gas phase as the volume of gas phase is known. The following formula can be used: pV = nRT, where
p – the pressure of the gas, N/m2 or hPa;
V – the volume of the gas, m3;
R – the universal gas constant, J/mol*K; R = 8.314 J/mol*K;
T – the absolute temperature of the gas, K;
n – the amount of the substance of gas (number of moles) [mol]; the number of moles n = 1 corresponds to 22.4 L gas under normal conditions.
A good example is Pereira et al, 2012 Article An OxiTop® protocol for screening plant material for its bio...
Anne Menert In the case where I only have pure methane (used as standard gas for GC), can i possibly calculate the 20% in air?
Which volume of bottles do you use? May be standard gas for GC would be too expensive to use as component of headspace. If you plan to make the gas mixture yourself, you need to make it by trial and error while testing the concentration of headspace. Far more easier and cheaper would be ordering the gas mixture needed.
Anne Menert i will be using 100 mL serum bottle. Unfortunately, that's the only CH4 available for me. Perhaps you can kindly share how to make the gas mixture?
There is a Book Gas Mixtures: Preparation and Control
Still it is better to obtain in advance prepared gas mixture to guarantee the correct ratio of gases (20% methane, 80% air). Good tips on cultivating methanotrophs can be found: Article Semrau JD, DiSpirito AA, Vuilleumier S.. Facultative methano...
Article Deciphering Community Structure of Methanotrophs Dwelling in...
Article Cultivation of Important Methanotrophs From Indian Rice Fields
It is also possible to use 0.5-1.0% methanol, as some methanotrophs are facultative methanotrophs being able to consume other substrates besides methane.- Badges
- Science method