I have Glycerol stock(6monts old) i need to prepare a working stock. what would be apropriate method for this.
should i streak over plates n culture them on broth or i can directly culture them on broth.?
Dear Bhim,
You can directly take 10-20 micro litre of sample and added it to 100 ml broth (nutrient broth or anyone in which you want to culture your isolate). First incubate it till it grows (approx. 48 h) and then sub-culture in new broth so that you can get only (and majorly) the active cells. This will be your working stock.
*Also you can check the purity of the culture by serial dilution spread plate technique (on basis of morphology, color, shape).
** Sub culturing is essential because in the first culturing all the cells are in in-active state due to preservation at very low temperature. Cells needs to be active. In 2nd culturing the active cells grows (divides) fastly and in exponential phase that is the necessity of any experiment.
Hope this helps. Thanks.
Ideally you should streak over plates and then inoculate individual colonies but you can use it to inoculate broth directly. Though in second option it may take some time for the culture to revive
yes i got it......
i streaked on plates, incubate, pick a isolated colony and inoculate into frsh broth.
thank you all.
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