For traditional cationic lipid reagents, we recommend testing various amounts of transfected DNA (0.25, 0.5, 0.75 and 1µg per well in a 24-well plate) at two charge ratios of lipid reagent to DNA (2:1 and 4:1; see Figure 5). This brief optimization can be performed using a transfection interval of 1 hour under serum-free conditions. One 24-well culture plate per reagent is required for the brief optimization with adherent cells (three replicates per DNA amount).
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