Nucleic Acids Extraction
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:Author:  Hailing Fang
:Created: 20240709
:Updated: 20240709

.. contents:: Contents
   :depth: 2

Genomic DNA Extraction from Animal Cells
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Genomic DNA Extraction with Phenol-Chloroform
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:Protocol ID: p0001

**Objective**

**Theory**

**Materials and Reagents**

*Reagents*

    Phenol;
    Chloroform;
    Isoamyl alcohol;
    TE buffer (Tris-EDTA);
    Proteinase K;
    RNase A;
    Centrifuge;
    Ethanol or isopropanol;

**Procedure**

1. Cell Lysis:
    Resuspend cells in a lysis buffer containing SDS and Proteinase K to digest proteins and lyse the cells.

2. Phenol-Chloroform Extraction:
    Add an equal volume of phenol:chloroform

3. alcohol (25:24:1).
    Vortex and centrifuge to separate phases.

4. Aqueous Phase Collection:
    Carefully collect the upper aqueous phase (containing DNA) without disturbing the interface.

5. Chloroform Extraction:
    Add an equal volume of chloroform, vortex, and centrifuge again to remove any remaining phenol.

6. DNA Precipitation:
    Add cold ethanol or isopropanol to the aqueous phase to precipitate the DNA. Incubate at -20°C for 30 minutes or overnight.

7. DNA Collection:
    Centrifuge to pellet the DNA, wash with 70% ethanol, and air dry the pellet.

8. DNA Resuspension:
    Resuspend the DNA pellet in TE buffer or water.


Total RNA Extraction from Cells
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