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7.

Isolation and cultivation of


microrganisms
Definition: culture medium, culture,
pure culture, mixed culture, isolates
Aseptic techniques and relevance to the
isolation and cultivation of microbes
Establishing a pure culture:
Sterilization (dry vs autoclave):
 dry heat : less effective, absence of water, kills by
oxidation of cell, denaturation of proteins. Eg :
direct flaming, hot air sterilization(items put into
ovens). Slow, not suitable for heat sensitive
materials.
 Autoclave : above 1000C to destroy endospore
(saturated steam under P). Usually 1210 C and 15
pound of P for 15 mins. Degrading nucleic acid.
Different streaking methods
Streaking in obtaining isolated colonies:
a typical colony is picked off and
streaked on a second plate, and
repeated until only one type of colony is
observed through several isolations –
single homogenous species.
Maintaining and preserving
pure cultures (stock cultures):
Agar slants vs petri dishes and liquid
medium
Preservation under oil
Freeze drying (lyophilization)
Storage in liquid nitrogen