1. DNA Extraction and Isolation
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DNA Extraction and Isolation

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DNA in the Cell
DNA is found in cells, particularly in the nucleus or most animal and plant cells.

4. Why Extract the DNA? Genetic testing Body identification
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Why Extract the DNA?
Genetic testing
Body identification
Forensic evidence analysis

5. How do we get the DNA from cells?
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How do we get the DNA from cells?
Extracting and Isolating DNA from cells take place in essentially 3 steps:
Additional step for plant cells: grind plant matter to break cell walls
Lysing (breaking) the cells to release DNA
Separating cellular and protein debris from DNA
Precipitating and isolating the DNA

6. Presentation Name
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Grinding Plant Matter
Plant cells contain cell walls which are tough and surround the cell membrane
Plant material should be physically ground so that the cells walls are broken down

7. Step 1: Lysing Cellular and Nuclear Membranes
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Step 1: Lysing Cellular and Nuclear Membranes
Both cell (plasma) and nuclear membranes are made of lipid (fat)-based molecules called phospholipids in 2 layers called a bilayer
Nuclear membrane is a double bilayer

8. Step 1: Lysing Cellular and Nuclear Membranes
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Step 1: Lysing Cellular and Nuclear Membranes
A lysis buffer which contains a mild detergents helps to break apart both the cell and nuclear membrane to release the DNA contents

9. Step 2: Separating Debris from DNA
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Step 2: Separating Debris from DNA
DNA is wrapped in a tightly coiled form called a chromosome
DNA wrap around proteins called histones so that the coiling is more compact

10. Step 2: Separating Debris from DNA
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Step 2: Separating Debris from DNA
Adding salts (and proteases, enzymes that break down proteins) helps to remove the histones and other protein debris by neutralizing the DNA’s charge

11. Step 3: Precipitating the DNA
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Step 3: Precipitating the DNA
The DNA is still dissolved in water at this point.
Adding cold alcohol (either ethanol or isopropanol) will make DNA less soluble in water and it will precipitate out in the alcohol layer (top layer)