I am looking for protocol or methodology to obtain single cell suspension of M. bovis BCG. Normally they occur in clumps when I see them under microscope. I found some methods like Sonication, passing though 27G needle, passing through 8 μm pore size membrane filter etc.
Does anyone have experience? Let me know which one works good to get relatively better single cell suspension of BCG.
Did you try to cultivate BCG in Dubos medium which contains Tween 80 and is adapted to Mycobacteria ? I suppose that Tween 80 could help you to obtain well dispersed bacteria after a slight sonication.
Hey Erin, Thanks for answer. I have already grown suspension of BCG bacteria in liquid culture. But mostly they are in clumps (observed under light microscope). Now how can I get single cell bacterial suspension?
Hello
i agree with Erin for the agar , in link , there are many protocols you can choose the best for you .
Hope help you
Good Luck
http://www.jimmunol.org/content/166/10/6367.full.pdf
http://www.nature.com/mi/journal/v6/n5/full/mi2012131a.html
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3071241
https://docs.google.com/viewer?url=patentimages.storage.googleapis.com/pdfs/US20130059364.pdf
http://onlinelibrary.wiley.com/doi/10.1046/j.1472-765x.2000.00619.x/pdf
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0002461
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC143408/
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