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        Titration of Lentiviral Vectors

        互联网

        1513

         

        Day 1 (9-12am)

        • plate 30.000 293t per well on 24 well plate in 1 ml medium/well

         

        Day 2 (4-6pm)

        • count cells on 1 well (should have 60-80.000)
        • transduce the cells with 6, 4-fold serial dilutions in 250� total vol; take 50� for non-conc. or 1� for conc. vector stock for as a first dilution

         

        Day 3 (9-12am)

        • add 1ml of medium

         

        Day 5

        • split cells and analyze fluorescence by FACS and read percentage from linear values (usually 5-10%) Titer is a number (percentage) of cells transduced by a given vol and counted on D2 e.g. 1� gives you 10% of positivie cells and you had 50.000 cells on D2, so you have 5.000TU/� --> 5x106 TU/ml

         

        Day 7-8

        • isolate DNA for QPCR

         

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