DNA extraction from plant material Flashcards
1
Q
What are the 4 steps of DNA extraction?
A
- Homogenisation
- Lysis
- Separation
- Precipitation
2
Q
What is homogenisation?
A
Plant sample ground using pestle + mortar
3
Q
Whats the reason for homogenisation?
A
To break open cell wall
4
Q
What is lysis?
A
Sample is mixed with detergent, salt, + protease enzymes
5
Q
Whats the reason for adding detergent?
A
Disrupts phospholipid bilayer of cell membrane, releasing DNA
6
Q
Whats the reason for adding salt?
A
Neutralises charge of phosphate groups - reduces solubility of DNA in water
7
Q
Whats the reason for adding protease enzymes?
A
They digest the histone proteins, just leaving DNA
8
Q
What is separation + precipitation?
A
Ice-cold ethanol added to mixture - causes DNA to be seen as white strands
9
Q
How is the DNA removed from the mixture?
A
Scooped out using inoculating loop
