Density gradient construction

Sphaeroplasts were prepared by slight modifications to published methods [12,13]. Lysis of sphaeroplasts was effected by a combination of osmotic lysis and gentle mechanical disruption [14]. Discontinuous sucrose-density gradients were constructed and fractions were then assayed for protein, PG and marker enzymes for different organelles. 

The approach initiated by Lee et al. [28], was rapidly adopted and as a consequence, several kinetic energy conjoint gradient correction functionals were generated. In order to compare the behavior of kinetic energy density functionals constructed in the context of LS-DFT and some members of the... 

The density was measured at room temperature using a density gradient column constructed from toluene and carbon tetrachloride. The column was prepared according to ASTM-D 1505 Method B. The column was calibrated with glass floats of known density. The accuracy of density measurements was estimated to be 0.001 g/cm Small pieces ( 25 mm ) were placed in the column and allowed to equilibrate before the measurements were taken. 

Figure 34. Zone convection electrofocumi of a 40% Ampholine solution, pH 4-6. The electrofocusing time was 5 days and the voltage 800-1200 V. The contents of two different compartments were taken and diluted to 0.5%. They were electrofocused by density gradient electrofocuaing method. Each fraction covered about 0.5 pH units. pH-gradients with specified shapes can also be constructed by adding or removing different fractions (Valmet, 53).

During the incubation, construct sucrose density gradients. Pour 1 ml of 60% sucrose solution into an autoclaved ultracentrifuge tube. Then, add 1 ml of 40% sucrose solution, and overlay with 8 ml of 30% sucrose solution. 

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