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lentivirus packaging with large insert


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#1 abubu

abubu

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Posted 24 July 2011 - 08:03 PM

Hi everyone
I am facing some problems with lentivirus transfection.
My problem is more severe though. The total length of the plasmid is about 11000bp, with a 3400bp insert. (OD260/280=1.82)
I used lipofectamine2000 for transfection. I got poor flurescence dectected either using 293T or 293FT as packageing cell line.The packageing vector we used is the 3rd generation system(REV,VSVG,pMDL).The transfection of PLL3.7 and Neg control(same backbone with no insert) seemed to be fine.
And I have done some optimization by changing the ratio of plasmid DNA and lipo2000, with disappointing results.
Is there any suggestions?




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