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Stages of DNA Isolation

- Homogenization


- Deproteinization


- Precipitation

3

Homogenization does?

Cell walls of the bacteria are broken down

Deproteinization does?

Proteins in the cells are removed?

Precipitation does?

DNA is concentrated by removing it from the aqueous solution

Homogenization components

- EDTA


- SDS


- Proteinase K


- Heat

4

Deproteinization components

- Proteinase K


- Phenol/ Chloroform

2

Precipitation components

- Sodium acetate


- Isopropanol

2

EDTA

Destabilizes the cell wall

SDS

Disrupts the cell membrane

Proteinase K

Degrades the proteins in the cell

Heat

Disrupts the cell further

Phenol/ Chloroform

Denatures and separates proteins from DNA

Sodium acetate

Neutralizes the charge on the DNA sugar (phosphate backbone) making it less hydrophilic

Isopropanol

Precipitates the DNA out of a solution