Sponge samples were collected approximately a month ago. The plan now is to separate bacterial cells from sponge cells as done by Thomas et al 2010. Sponge samples available are in a) 20% glycerol b) RNA later and c) plain, i.e. straight into the freezer. Glycerol and RNA later stocks are at -80C, whilst sponge tissue alone was stored at -20C. Which sample would have the highest number of intact bacterial cells?
I would assume your best sample would be the one in glycerol. (Glycerol being able to protect the bacteria from some damage during freezing.) But I have never compared the results myself.
Maybe the following article/paper would be helpful? They talk about freezing bacteria in a variety of ways and survival/culturability.
Survival of specific strains is difficult to predict. Generally glycerol is accepted as a very good protector. What you have to realize is however that by culturing you will only find (i) the survivors, i.e. the most resilient strains/species and (ii) the culturable cells under the conditions you chose, e.g. medium, temperature, (an)aerobic, .... It is in (ii) that the real problem resides.
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