Investigating organelles

Cell fractionation
1. Cold
  1. To reduce enzyme activity which might break down organelles
2. Isotonic
  1. Prevent bursting/shooting by osmostic loss/gain
3. Buffered
  1. To maintain constand pH
Homogenation
1. Cells broken up by homogeniser (blender)
  1. Resultant fluid (homogenate) filtered to remove complete cells and large debris
Ultracentrifugation
1. Tubes of homogenate spun at high speed
2. Tube of filtrate placed in ultracentrifuge and spun at low speed
  1. Heaviest organelle forced to bottom (sediment)
    1. Fluid at top (supernatant) removed leaving only sediment
      1. Supernatant transferred to another tube and spun again at higher speed
        Next heaviest organelles forved to bottom- process continued with increase in speed each time

Resource summary

	Created by hannah.lindridge almost 11 years ago