After 5 minutes, the mixture should come together and a film should form on the bottom of the pan.
The butter will melt and the sugar will dissolve, and over the course of the next 10-15 minuthe the mixture will gradually heat up and thicken.
Add the saffron and wine, and whisk continuously over the steam for about 20 minutes; the mixture will lighten in colour and texture as the eggs cook.
Cells were collected, mouse macrophages were lysed by adding 0.05% SDS, and after 5 minutes the mixture was centrifuged at 1000 g for 10 minutes.
After denaturing at 75°C for 10 minutes, the mixture was maintained at 37°C for 10 min. MagPrep® Streptavidin Beads (Novagen) were added to the mixture to remove the hybrids and the free probes.
Briefly, 3 ug of plasmid was added to a 2.5 M calcium solution (Clonetech), mixed with 2 X HeBS (Clonetech) and after 20 minutes, the mixture was added to the cells.
After cool-ing for 10 minutes the mixture was centrifuged for 10 minutes at 10,000 x g.
After incubation at 30°C for 15 minutes, the mixture was boiled with DNS for 5 minutes and the absorbance was measured at 520 nm [ 43].
After incubation at 30°C for 60 minutes, the mixture was boiled with DNS for 5 minutes and the absorbance was measured at 520 nm.
