Figure 3.

Design of a 96-well lentiviral vector transduction assay using an in situ DNA extraction method. (a) Schematic of the method. Assay cells seeded in 96-well microtiter plates are transduced with serial dilutions of vector (75 µl/well final volume). Following an overnight incubation, cells are lysed by the addition of 25 µl of lysis buffer containing detergents and proteinase K (100 µl total cell lysate). Plates are then sealed and incubated at the indicated temperature hold steps to degrade proteins and denature the DNA, after which samples are analyzed by qPCR (9 µl sample/rxn) to detect vector sequence. (b) C_t values (mean and SD) of the seven-point qPCR standard curve (n = 3 qPCR replicates; symbol size was larger than error bars).