Cell Fractioning

Question 1

explain the process of cell fraction

Answer 1

1) Prepare cold, buffered , isotonic solution
2) mix solution with cell tissue and place in a blender (homogenise it )
3) filter mixture then spin homogenate at low speed using centrifuge
4) collect pellets formed and return supernatant to centrifuge to spin at higher speed
5) continue until all organelles required are isolated.

Question 2

Why does the solution have to be cold?

Answer 2

• reauces enzyme activity that breaks down organelles

Question 3

why should the solution be isotonic?
(same water potential)

Answer 3

stops water flowing in and out of cells

• hypotonic: (more water) organelles would burst due to osmosis
  -Hypertonic : (less water) the organelles would shrivel up due to water moving out of them by osmosi

Question 4

Why should the solution be buffered? (constant ph)

Answer 4

so that organelles (proteins) don’t denature or are damaged

Question 5

What is the order of organelles coming
out during cell fractioning? (centrifugation)

Answer 5

• nuclei
•chloroplast ( plant)
• Mitochondria
• lysosomes
•endoplasmic reticulum
• Ribosomes.

Question 6

Why do the organelles separate in that specific order?

Answer 6

they go from biggest (heaviest) to smallest (lightest)