Hi Priscila, first of all, in our lab we use Insect Xpress Medium (Lonza), supplemented with Pen/Strep, fungizide Partricine (you might try fungizone) and Pluconic (Gibco). We are doing them in round (not Erlenmeyer) shaking flasks (40 rpm) at 27 deg Celsius. Sf9 cells usually stop growing when the split-rate is too high (after weeks, they recover anyway). You should count the cells in a hematozyte counter and then split them at density of ~8-12 million/ml (in our lab, it takes about 3-4 days) to not less than 1 million/ml. When you infect the cells, 2 million/ml is a good cell density. We do the shaking for virus amplification, protein expression and cell maintanance. Try it, and if you still have questions, :-) Niko
Dr. Nikola Wenta Research Fellow D82 School of Biomedical Sciences University of Nottingham Queen's Medical Centre Nottingham NG7 2UH Tel: +44-(0)115-8230298 This message has been checked for viruses but the contents of an attachment may still contain software viruses which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation._______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
