in this PDF we will discover - What is organic extraction -Uses of the isolated DNA - Steps of the phenol-chloroform method - Disadvantages of this method - overall view

1. DNA organic extraction
Phenol-chloroform method

2. Organic Extraction
• liquid-liquid extraction
• most commonly used methods in molecular biology
• Phenol-chloroform-isoamyl (PCI) alcohol extraction method based on:
❑the differential solubilities of biomolecules in different immiscible
liquids.

3. By this Method we can isolate DNA from :
• blood cells
• muscle
• brain
• flower
• hair follicles
• dental pulp tissue
• epithelial cells
• osteocytes of bones

4. the isolated DNA can be used in :
gene
amplifcation
fingerprinting
single nucleotide
polymorphism
(SNP) studies
RFLP
gene
quantifcation

5. cell lysis
2 phases
DNA
precipitation
STEPS

6. Take care of PH of phenol

7. Disadvantage of this method
uses different
dangerous organic
solvents and hence
should be carefully
handled
laborious and time-
consuming compared
to the other methods
available

8. the pH of the phenol must be
kept slightly alkaline (pH 8)
because the alkaline pH keeps
the nucleic acids (DNA and
RNA) negatively charged, thus
allowing the portioning of both
into the aqueous phase