DNA Extractions
& Analysis
Author:
Nabeel Ahmed
Introduction
‘DNA extractions is a process in which we separates DNA from proteins,
carbohydrates and other chemical structures,
While DNA extraction methods have been around for over 150 years First
DNA .Since the first DNA extraction performed by Friedrich Miescher in 1869,
scientists have made extraordinary progress in designing extraction methods
that are more reliable, easier and faster to perform, more cost-effective and
produce a higher yield.
Type of DNA Extractions
Inorganic DNA Extraction
The principle is based on the high affinity of the negatively charged DNA and
RNA molecules to the positively charged silica coated on the magnetic particles.
Under high salt and optimal pH, the DNA or RNA molecules are tightly bound on
the silica surface of the particles.
DNA
Organic DNA Extractions
This is also known as Phenol: Chloroform Extraction Protocol. Organic extractions or Phenol–
chloroform extraction is a liquid-liquid extraction technique in biochemistry and molecular biology
for purifying nucleic acids and eliminating proteins. In brief, aqueous samples are mixed with equal
volumes of a phenol: chloroform mixture. After mixing, the mixture is centrifuged and two distinct
phases are formed, because the phenol: chloroform mixture is immiscible with water. The aqueous
phase is on top because it is less dense than the organic phase (phenol: chloroform). The proteins
will partition into the lower organic phase while the nucleic acids (as well as other contaminants
such as salts, sugars, etc.) remain in the upper aqueous phase. The upper aqueous phase is
pipetted off and care is taken to avoid pipetting any of the organic phase or material at the
interface. This procedure is often performed multiple times to increase the purity of the DNA.
ADVANTAGES AND DISADVANTAGES:
Its other main advantage is the fact that it can be used on a wide range of samples.
However, this method does also have some disadvantages including being very labor
intensive, being easily contaminated and exposing the scientist carrying out the
extraction to dangerous chemicals.
DNA
Methods & Materials
DNA
Reagents
• 1Ether (optional)
• Phenol: Chloroform (1:1)
• Tris EDTA (pH 7.8) (optional)
• M sodium acetate pH 5.2 or 5 M
ammonium acetate
• 100% ethanol
• Proteinase K
Equipment's
Centrifuge Micropipettes Micropipes Tips
Machine
Micropipes Tips Box Water Bath EPHEN DROP 1.5ML
DNA
Cell Lysis
Cell lysis is a process of breakdown of cell plasma membrane by
using of chemicals or by physical means depend upon the origin of
samples . We can be use a buffer solutions ,EDTA or viruses . But
during DNA extraction we use EDTA ,buffer solutions and centrifuge
machine for cell lysis .Buffer Solution is also use to maintain PH of
solutions .
Removal of Membrane Lipids
After Cell lysis the membrane lipids are
removed.
DNA
DNA Repair Mechanisms
DNA repair mechanisms are cellular processes that correct DNA
damage, ensuring its integrity and preventing mutations. These
mechanisms include base excision repair, nucleotide excision
repair, and mismatch repair, among others.
DNA
DNA Recombination
Understanding the process of DNA
0
recombination and its role in genetic
1
diversity and evolution
Recombination events, such as
0
homologous recombination and site-
2
specific recombination
Conclusion
In conclusion, DNA, or deoxyribonucleic
acid, is a fundamental molecule in living
organisms, carrying genetic information
that determines traits and characteristics.
Its structure, sequencing, mutations, and
repair mechanisms play crucial roles in
genetics and biology.
DNA
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