Well im doing a small study, to determine the efficiency of two different methods of surface sterillization methods on wheat plants, treated briefly with a M. anispoliae spore suspension. I so far found, that the methods i have been testing are insufficient at removing surface contaminats (or atleast all the spores from the suspension). Method 1 is subermerging the plantparts for 2 minutes in 70% ethanol, then 2-3 % sodium hypochlorite, then three waterbaths oh autoclaved ionized water. Method two is quite similar, but with significantly shorter times in each step (aproxematley 10-15 seconds), but with a 5-7,5 % sodiumhypochlorite concentration (this has been used for insect surface sterillization in the past). As i said, non of them seem to work, and thus im testing some tweaks to these methods. The first thing i tried, is adding 0,05% tween 20 to the hypochlorite, but i observed something peculiar. Ordinarily, when i submerge the plant parts into the hypochlorite, the part skipps across the liquids' surface, but when i submerge them in the dilution added tween, t does nothing. Also, the dilution has a strange, cloudy aperance. Im still waiting for the results of this test (incubating the sterillized plants on selective media, so it'll take a few weeks before i have anything conlcusive). with that said, here's my question. Do you think that the adittion of tween somehow ruins the hypochlorite?
I made the sodium hypochlorite dilutions twelve hours in advance (with the tween)
The goal of the study is to determine wheter the methods are sufficient at removing known surface contaminants (M. anisopliae)