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Chromosome Separation Attempt 2 (2/25/10)
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==Isolation of Chromosomes== 2 tubes of cells were extracted and used for this procedure. After being spun down, the palettes formed by the cells were very small and easily disturbed. After removing the majority of the supernatant, the cells tended to re-suspend in the remaining fluid, necessitating another spin in the centrifuge to re-form the palette. After this second spin the remainder of the supernatant could be pipetted out safely. After 1mL of the isolation buffer was added to each tube, the cells were easily resuspended via a few flicks to each tube. While the tubes sat at room temperature, the Triton X-100 filtration was performed. The X-100 filtration: The filter was added to the syringe first, the plunger was removed from the syringe, 500uL Triton X-100 solution was added to the syringe, the X-100 was then filtered (rather forcefully) *Each attempt to push the X-100 solution through the filter caused a large snapping sound, presumably from the filter *500uL was taken from the stock bottle, and only 100uL filtered out of the syringe *The solution is highly viscous, causing troubles when it was pipetted into the syringe and then to each tube The samples were allowed to sit at room temperature for 10 minutes. One sample was mixed via repeat-pipetting, using a syringe with a 22 gauge needle attached. The solution was pipetted 20x The second sample was mixed using the PowerGen125 Homogenizer for ~60 seconds.
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