Thank you all for your suggestions..... Have already started trying out some.
cheers
P.
-----Original Message-----
From: [email protected] on behalf of Nikola Wenta
Sent: Mon 22/02/2010 17:41
To: [email protected]
Subject: RE: insect cell line problems
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
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