Transfection - (Mar/18/2005 )

# Should I always split my cells at their mid log phase? Or does it only matter at transfection? How do I know that my cells are in the mid log phase or not too long in the plateau?

# Where should I prepare the media? In the laminar flow hood or on an opened table (risks for contamination?) ?

#If medium with serum contains trypsin inhibitor. What should I use to dislodge my cells?

#Should I always have HCO3- in the medium and what for?




Thanks.

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cells that are up to 50 % confluency and less than 80% are in their mid log phase



media must be prepared in the laminar flow

You should wash your cells twice with PBS. you may dilute trypsin in PBS. Inactivate the trypsin by adding medium.



usually, cells are in 5%CO2 air. This ensure a combination with the media to automatically form HCO3-. This ions give the pH stability and it's essential (but automatic) to have it in the media

Questions answered?

Fred

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