cell fractionation and ultercentrifugation

Question 1

conditions that solution needs to be in

Answer 1

cold

isotonic

buffered

Question 2

buffered

Answer 2

ph buffer

prevents damage to organelles

Question 3

isotonic

Answer 3

organelles should be the same water potential as solution to prevent osmosis causing organelles to shrivel or burst

Question 4

cold

Answer 4

reduces enzyme activity

when the cell is broken, enzymes are released which could damage organelles

Question 5

step 1 homogenisation

Answer 5

cell must be broken/opened using a blender

solution must be cold, isotonic and buffered

Question 6

step 2 ultracentrifugation

Answer 6

filtered solution is spun at high speed in centrifuge

separates the organelles depending on the density

Question 7

differential centrifugation

Answer 7

centrifuge spins at a high speed

centrifugal force causes pellets of nose dense organelle to form at the bottom of the tube

process is repeated at increasingly fast speed, removing supernatant and leaving behind pellets