BTN315: Practical Prep (2, part 1)

DNA = deoxyribonucleic acid

• Structure: double helix

• Components: A, T, C, G
• Base pairing: Adenine (paired up with) Thymine; Cytosine (paired up with) Guanine
• Replication: Semi-conservative
• Packaging: Tightly coiled around histones
• Found(in humans): Cell nucleus, mitochondria
• Charge: Negative

DNA ISOLATION

Process: Purifies DNA by removing cellular debris/contaminants (proteins, enzymes, organelles

etc)

1) Define lysis:
Lysis means to separate or break open.

2) Define a buffer:
A solution that provides an optimal environment. It resists pH changes when an acid or
base is added into a solution/stabilizes pH levels.

3) Why is lysis important in DNA isolation?

DNA is contained within cells (within the nucleus of a cell and the mitochondria). In
order to extract the DNA, we need to open the cell. Lysis allows us to do this.

4) Mention 2 important components of a lysis buffer

Detergent and Proteinase

5) Provide the function of the 2 components of the lysis buffer

a) Detergent – disrupts lipid bilayer of the cell membrane, causing the cell to break
open/lyse and release all of its cellular contents.
b) Proteinase – enzymes that break down and digest proteins/helps to overcome
protein-protein interactions.

6) Mention common detergents and proteinases used in the lysis buffers

a) Detergents: SDS and Triton X-100
b) Proteinase K