Methods I

Question 0

Cell Growth

Answer 0

-Unlimited number of cell divisions = Immortal cell
-Most are limited and maintained by Telomerase (DNA & Proteins)
>a way to keep track of cell generation and count divisions
-Cell Cycle Checkpoints can also limit #of divisions
>can be overcome by *Oncogenes (cancer genes)
>can be introduced or induced (gene transformation)
-A Clonal Population is derived from one cell

Question 1

Cell Isolation

Answer 1

• Collagenase: used to break down collagen which holds many cells together
• EDTA: can get rid of calcium w.o using acids *Chelates (v.)
• FACS: makes cell of interest fluorescent
• Laser beam can cut out region of interest
• many many more

Question 2

Heterocaryon

Answer 2

• 2 cells can be fused into one (2 separate nuclei)
• Exhibits properties of Both cells
• Membranes will float around and mix together
• Cells don’t like to have multiple nuclei

Question 3

Monoclonal -Antibodies

Answer 3

-Fusion of “X-antibody” and tumor cells (divide forever)
-Only hybrids will grow on selective medium
-Shake
-Allow cells to multiply and test the “sup”
=Creates Clonal Culture
*Polyclonal cells are more recommended

Question 4

Cell Fractionation

Answer 4

1. Osmotic Shock:
2. Sonication:
3. Smush through small hole:
4. Blender
5. Centrifuge
6. Velocity Sedimentation
7. Equilibrium Sedimentation

Question 5

Column Chromatography

Answer 5

• Ion Exchange: Charge
• Hydrophobic: hydrophobicity
• Gel Filtration: Size
• Affinity: ability to bind
• HPLC-high pressure used to speed up column excretion
• Salt can be added to the column to break bonds allowing more stuff to come out