I have been trying to isolate single colony units from what I believe to be mixed culture. I have tried a few different recipes, LB, Nutrient Broth, 523, YES, TSA10, KB, Ammonia Nitrate...
AN agar limited the growth substantially but still no single colony units are forming.
Almost every time I think I have a pure culture and do DNA extractions, send to lab, and I get mixed results for different genes from the same sample when I blast the results so I want to go back to my deep freeze cultures and start again trying to get colony forming units.
several isolates from different sources have the same milky aura, others have a more holographic slime, but for the most part the streak plates never form single colonies. I have done a few tests for nitrogen, nitrate, and ammonia, and the most interesting ones I have found reduce nitrate and also solubilize phosphate media quickly. Some of the cultures are definitely bacillus, others I'm not so sure. I have a book of culture media for environmental biology but it doesn't have a really nice key for finding appropriate media. Any opinions on what to do to get single colonies would be highly appreciated!
Both viscous and viscoelastic media alter the motility of flagellated bacteria without affecting their viability. Significant cell death occurs at high glycerol level but not in PEG/PEO media which also causes loss motility due to increase shear viscosity, but the majority of immobilized bacteria remain viable. .For further details consult https://bmcmicrobiol.biomedcentra.com
April Micci,
Motility detection media with tryptose and agar ; magnesium chloride-malachite green enrichment broth ; semi solid nutrient agar; motility test media with and without triphenyltetrazoliumchloride (TTC) for gram positive and gram negative motile bacteria; Lactose TTC with Tergitol; ENDO Agar for microbiology; McConkey's Agar with crystal violet and 0.15% Bile salt; Salmonella Chromogen Agar for microbiology; Malachite Green Broth of Microbiology etc can be of help to you both for detection as well as cultivation of the motile bacteria.
With regards,
Dr. Abhishek Mukherjee
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