I wanted to perform a quick experiment (10-25 minutes) which will help me to check microbial presence in soil, I have found many culture techniques but those at least take 8 hours of duration, I wanted to know fastest and reliable methods just to isolate bacterial presence in a given soil sample. Identification of type of bacteria is not necessary.
Best Luck.
Article Principal methods for isolation and identification of soil m...
Dear Shivam,
Though most analyses of soil microbial communities require more time and effort, you should be able to verify the presence of bacteria in samples by simple microscopy in which bacterial spores and cells can be visualised. Clearly spherical, rod and spiral-shaped objects may not in fact be bacteria, but if you are careful about determining size ranges you probably won't confuse them with other material including fungal spores, etc.
Regards, Andrew.
You can check bacterial abundance with DAPI staining techniques, viability with Live/Dead stain and INT and enzymatic activity by FDA hydrolysis.
Gram stain! Soak soil sample in distilled water for like 10 mins, filter and then use a drop of the filtrate to prepare your slide for Gram stain. View under microscope! All the best
The time is too short to actually ascertain the accuracy of result but i think molecular methods would prove more reliable in this case.
Best luck
Which molecular methods? Can you be more specific about them?
Thanks
Shivam
Molecular methods like DGGE or next gen sequencing (e.g. Illumina) are better but not quick or cheap. The best bet is microscopy with some stain ( DAPI, Gram or acridine orange for example) but it may not be easy to see many cells due to the loe cell density. Pre-culturing and micrscopy or substrate induced respiration measurement (CO2 evolution) will tell you there is something there, but wont tell you what is there or in what proportions each kingdom/genera/species/kingdom is in the original soil.
You're right Tracy but you still have to sequence the 16S fragments and BLAST which will take longer
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