I'm planning to subculture bacteria in blood agar plates and store them in blood agar broths
Can this be done?
First of all you will not succeed in making such broth. You may be able to make a serum broth but seldom blood medium broth. Secondly it is never a good medium for long term preservation. It is mostly used for maintaining working culture that too by frequent passaging at short time intervals.
It may be possible to store some cultures of bacterial on blood agar slants for a month or two. After the culture has been established on the slant, cover the growth with sterile mineral oil and refrigerate. You may subculture directly from the oil covered slant with a sterile loop, and place the culture back in the refrigerator
http://generalbacteriology.weebly.com/culture-media.html
Check -Media used for storing the bacteria for a long period of time. Examples: Egg saline medium, chalk cooked meat broth.
Dear Sony MAc
You can't store your bacteria for long period of time in broth, So after subculture in blood agar broth, incubate overnight then transfer the 500 microlitre of over night incubated culture into 2microlitre cryovial or screw top tube then add 500 microlitre of 50% glycerol solution. Mix it gently, Freeze the glycerol stock tube at -80°C. The stock is now stable for years, as long as it is kept at -80°C. Subsequent freeze and thaw cycles reduce shelf life. To recover bacteria from your glycerol stock, open the tube and use a sterile loop, toothpick or pipette tip to scrape some of the frozen bacteria off of the top. Grow your bacteria overnight at the appropriate temperature.
I would recommend using a more "standard" broth for storage, like LB or TSB, with 25% glycerol. Store them at -80°C. I do this all the time with some very fastidious, difficult to culture anaerobic gram negative species (Tannerella forsythia and Porphyromonas gingivalis) which usually grow on a kind of blood agar, and they grow just fine after I remove them from storage and subculture them.
If you are not sure this will work, I would recommend testing this by keeping your initial culture alive by subculturing, and freezing some of it, then plating the frozen stock back out to test its viability before you discard your cultures. Good luck with your culturing.
P.S. A note on freezing cultures: Do NOT defrost the whole vial of frozen bacteria. Just scrape some of the frozen culture from the storage vial and plate the ice onto your agar. I repeat DO NOT defrost the whole storage vial.
Dear Researcher,
just as Said Amrani and Richard R Facklam said earlier and thats what have being on my mind, why dont you just use the culture directly on blood agar slants and then subsequently store at cooler temperatures, because what you are proposing is a bit futile.
wishing you well in your research
Agar slants don't last very long. Freezing is the only way to achieve long term storage.
Blood agar cannot be broth! It is a solid medium when agar is there. Brain Heart infusion broth or Trypticase soy broth with 15-20% glycerol is a good storage medium for bacterial stock. Or you may stock it in Blood agar slant at 4 degree C for limited period. Good luck
i agree with Jalaluddin above- use Heart Infusion Broth with 15%glycerol- super for long term storage and recovery works well for fastidious bacteria. used in Hospitals.
Acually, many moicrobiology labs, particularly in developing countries may not have -80 degree C biomedical refregerator, so in this situation making slants, kept in regular Refrigerator at -4 degree C and re-subculturing every 3 months is the appropriate method. I agree with all collages that the bood agar or broth medium is not a wise choice to store your bugs for a long period.
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