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Procedure
Plating Pour plate technique · Melt sterile plate count agar medium by boiling. Cool the molten agar and maintain in a water bath at 45°C - 47°C. · Label the petridishes at the bottom indicating the dilution, sample details etc. using a glass marker · From the dilutions transfer 1 ml of inoculum into petri dishes (two dishes for each chosen dilution) · Pour approximately 20 ml of molten, tempered agar into each of the petri dishes and mix with the inoculum by swirling the plates a few times. · Allow the plates for sometime to solidify the agar and incubate at appropriate temperature in inverted position. · After incubating for 24 h or 48 h observe for colonies in each dilution and record the results. · Look for the gradual reduction in the crowding of colonies with increased dilution. · At some higher dilution, the colonies would be well distributed so that each colony could be isolated. Each colony has grown from a single bacterium, thus this method helps in purifying mixed cultures of bacteria. |