... why baculovirus at all (unless you happen to receive the working system from a colleague:) ? there is an efficient transient transfection protocol with PEI. Much easier, faster and fewer variables. Yield excellent as well. Only one "pet" per protein expression trial ;) ( Methods Mol Biol. 2009;498:199-227. High-throughput insect cell protein expression applications.
Buchs M, Kim E, Pouliquen Y, Sachs M, Geisse S, Mahnke M, Hunt I. Biologics Center, Novartis Institutes for Biomedical Research, Basel, Switzerland. ) cheers, David 2010/2/22 Carmen P Pena Diaz <c.p.pena-d...@2006.hull.ac.uk>: > > Dear All: > > We recently started tissue culture work with SF9 insect cell line for protein > expression. > > Since we obtained the cell line we started culturing in TC100 insect media > and then shifted to Grace's supplemented insect media. After the passage the > cells failed to continue growth and that was almost 3 weeks ago. We're not > keeping the cultures with antibiotics in order to help them grow better. > > My boss obtained a second batch of cells from the firm and put them to grow > directly in Grace's media, in case the media changing might have affected the > cells irreversibly. Nevertheless, they're still not growing properly. > > On the other hand, we also have many cells not attaching to the surface as > expected. Since this is actually needed to calculate baculovirus titer, I > have not started trasnfections due to these problems. > > Anyone can suggest/comment on my cell culture problem? > > > thank you very much > > > Priscila > > > > ***************************************************************************************** > To view the terms under which this email is distributed, please go to > http://www.hull.ac.uk/legal/email_disclaimer.html > ***************************************************************************************** > _______________________________________________ > Methods mailing list > Methods@net.bio.net > http://www.bio.net/biomail/listinfo/methods > -- David Minde MSc (TUM) Cellular Protein Chemistry Room 707 Department of Chemistry Faculty of Science Utrecht University Krytgebouw Padualaan 8 NL-3584 CH Utrecht The Netherlands office phone +31 30 253 4105 (magic:) android +31 652614443 home: Griftkade 4 bis 3572 TW Utrecht Science is what happens while we are making other plans (~John Lennon) _______________________________________________ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
