I want to extract microorganism from specific soil and then inoculate them into another sterilize soil. Does anyone know how to extract microorganism from soil? Is there any standard methods? I am wondering if I can use distilled water to extract microbes? How long should I shake the soil solutions and how much solutions should I use to inoculate into sterilize soil?
I wonder "extract" would be a correct word. You can use the word "seperate" or isolate".
Now in order study microbe from soil we usually do serial dilution, where we take 1:10 ratio of soil: sterile distilled water, vortex it and then plate usually 0.1ml. But this protocol is usually used to perfom SPC rather than to isolate microbes from soil (though SPC cannot be done unless you have all the microbes)
Now the protocol is not standerdized and hence you can change your soil: sterile distilled water as per your convinience (depending on your idea about number of microbes present in soil), dilution numbers and even plating volume. Vortexing time is also usually not fix but its usually shake it till you break the soil.
The above mentioned protocol could be used to seperate your microbe from the soil, tranfer in to sterile distilled water (diluted or undiluted) and use that distilled water to inoculate in to sterilize soil. Here you will tranfer small amount of distilled water containg your soil microbes in to your sterlize soil (presumbaly if you are using sterlize soil agar plate) and hence you might miss some microbes. But with this procedure you can inoculate many plates.
Another way is to filter your "soil tranfered distilled water" and use the filter to inoculate your plates. In this method most of the microbes would be attached to filtered and chances are that they would get isolated. But you can use one only plate per filter
You can also finely crush your soil and use the soil directly.
I am not sure wether you are using sterlize soil agar plate or liquid broth but above method can be used for broth also.
You can also use interesting method like diffusion chamber or microbial trap to directly use the insitu environment (natural soil) for isolation of soil microbes. People have isolated more microbes than the above mentioned methods. I am putting a link of paper which review such method. Hope it helps
http://www.sciencedirect.com/science/article/pii/S1369527413001458
Isolation of soil microbes could be done by collecting the soil samples from desired location and performing serial dilutions of the sample (1:10 ratio).
Then you can go for pour plate method or spread plate method for knowing cfu of the soil.
further you can select the bacteria (in which you are interested) and get pure culture of the isolate by streaking on agar plates
after getting purified culture of the bacteria you can add the grown culture in the sterlized soil ( but before that u have to check water holding capacity of the soil) so that you can add appropriate amount of culture.
U can get help from following linkhttps://www.jove.com/science-education/10099/culturing-and-enumerating-bacteria-from-soil-samples
Dear Yuan Liu
Pls. find the attached files
http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2011.02209.x/full
https://www.cabdirect.org/cabdirect/abstract/20013064240
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC140122/
http://www.cellulosechemtechnol.ro/pdf/CCT3-4(2012)/p.227-242.pdf
http://link.springer.com/article/10.1007/s12223-012-0179-5
http://www.sciencedirect.com/science/article/pii/S0038071797002320
https://www.ncbi.nlm.nih.gov/pubmed/21221926
Hoping this will be helpful
Regards
Prof. Houda Kawas
As per the suggestions of other researchers you should go for serial dilution.
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