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The cultures were further incubated for 24 hours. Day 2 post-culture treatment (presence and absence of S9 cultures) Cell count was performed and the
suspension growth (SG) and relative suspension growth (RSG) were calculated for each culture. SG) for 2 days expression period
were calculated and SG of each test cultures were compared to control. SG (mean control SG) X 100 Based on the RSG value obtained the concentrations
chosen for the plating (viability assessment and mutant frequency) includes at least one dose level with an RSG value of 10-20% a no effect dose and a minimum of two further doses between this range of concentrations. CM10 media was prepared in sterile universal bottles. The procedure was done under subdued light due to TFT sensitivity to light.
The cells were counted and 2 x 104 cells were transferred onto microscope slides followed by centrifugation (cytospin at 450 rpm for 5 minute). Y in kratom extract info phosphate buffer) for 5 seconds. The excess stain was then
drained onto absorbent paper and the slides were transferred into basic solution dye (methylene blue in phosphate buffer) for another 5 seconds. Finally the slides were rinsed Kratom To Suboxone Livingston briefly in the buffered water (pH kratom extraction methanol 7. The slides were mounted with DPX and microscopic examination was then ultra premium kratom carried out similarly as described for WrightGiemsa staining procedure. AAD double staining for apoptosis detection In principle the cell membrane of live cells is covered by phospholipids (lipid bilayer) in which phosphatidylserine is located on the inner layer of the plasma membrane.
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