A
B
Figure 4. Optimisation experiments of Transfectam to DNA Ratio in transfected cells
. Duplicate independent transfections were carried out using the carrier to DNA ratios indicated below. One set was analysed by slot blot hybridisation (A) using total DNA (4 m
g DNA/slot), and the second set (B) by dot-blot hybridisation, using chromosomal-enriched DNA (1.0 m
g DNA /blot) from the Hirt fraction described under Materials and Methods. In both cases, control lanes were: C+
=
pCMVL DNA (109 copies = 1 ng), and C -
=
CHSE-214 cell DNA (1.0 m
g). Boxes 1 to 6 represent different Transfectam/DNA charge ratios expressed in m
g, and R =
the corresponding ratio. (Box 1) 2.5/5.0, R=0.5; (Box 2) 2.5/2.5, R=1.0; (Box 3) 2.5/1.0, R=2.5; (Box 4) 6.25 /1.0, R=6.25; (Box 5) 6.25/5.0, R=1.25; (Box 6) 12.5/10.0, R=1.25. The vector pCMVL DIG-labelled was used as probe.