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Wednesday, July 16, 2008

SHAKER FLASK

AIM:
To inoculate the shaker flasks with the preserved freeze culture of Pichia pastoris
PROCEDURE:
1. The prepared media was autoclaved.
2. Inoculated the autoclaved and cooled shake flasks media with the provided pre-inoculum culture under sterile conditions. Usually 4-5 % of the shake flask volume was inoculated into it.
3. Incubated the inoculated flasks on a shaker with the RPM and Temperature set suitable for the growth of the P.pastoris. Here, Orbital Shaker was used, with the RPM set at 200, and temperature at 30 oC.
4. Allowed the culture to grow for 24hours, before inoculating to the next fermenter, so that P.pastoris has grown to a suitable level.