Acetone And Methanol Lab Report

Chloroform, Acetone and Methanol) for sequential extraction. First of all plant materials were soaked in Hexane for 2-3 days with regular stirring. The mixture was then filtered by using Whatman No. 4 filter paper in to a glass beaker. 300ml of other solvent, Chloroform, was then added in the residue for 2-3 days. Similarly the mixture was filtered by using whatman No. 4 filter paper. After chloroform, Acetone and Methanol were added in residue for sequential extraction. Solvents; Hexane, Chloroform, Acetone and Methanol were evaporated by placing the extracts in fume hood for 7-10 days.
Stock solution prepration:
To prepare 40 µg/µl stock concentration in Dimethyl sulfoxide (DMSO), 40mg of each plant extract was suspended in DMSO, inside Laminar flow hood. Stock solutions were filtered through 0.22 µm filter and then stored at -20ᵒC.
Half an hour before use, medium, trypsin EDTA, and 1X PBS were incubated at 37ᵒC. The previous medium of the cultured cells was removed from the flask. Flask was washed with the 5 ml of 1X PBS. One ml of trypsin EDTA was added in the flask, and meanwhile placed in the incubator for 3-5 minutes, to detach the cells from the walls of flask. To normalize the trypsin EDTA, 5 ml of fresh media was added to the flask, and cells were shifted to sterilize culture tubes, and spun for 2 minutes at 2000 rpm. The cell pellet then obtained was re suspended in 10 ml of DMEM with antibiotics, into single cell suspension. (5-10
Approximately 2 x 104, 5 x 104, 2 x 10 5 and 5 x 105 cells were seeded in 96, 24, 12, and 6 wells tissue culture plates respectively, and incubated at 37ᵒC in 5% CO2 for 24 h, preceding cell viability assay or