DNA Extraction of

White Onion
Group 5
Gene

● Basic physical and

functional unit of
heredity.
● Composed of DNA
● Arranged on
chromosomes
Histone

● Provides structural
support to
chromosomes.
Nucleosomes

● built from histone

proteins around
which DNA is
coiled.
Nucleotide ● Monomer of Nucleic Acid
● Has 3 parts
● DNA bases - A,C,G,T
● RNA bases - A,C,G,U
Structural difference of DNA and RNA
Structural difference of DNA and RNA
 Different Bases of DNA and RNA

DNA Adenine and thymine - (A)pple on the (T)ree

Guanine and cytosine - (C)ar in the (G)arage

RNA Adenine and Uracil - (A)pple (U)nder the tree

Guanine and cytosine - (C)ar in the (G)arage

How bases are paired
Secondary structure of DNA
Diphenylamine
The reaction between the reagent and deoxypentose from DNA results in the
development of a blue color. Acidic conditions convert deoxyribose to a molecule
that binds with diphenylamine to form a blue complex. The intensity of the blue
color is proportional to the concentration of DNA. The reagent does not react with
the ribose sugar in RNA and does not form a blue-colored complex.
 Diphenylamine Test
Purpose - To detect the presence of Deoxyribose
 Diphenylamine Test

Components of Diphenylamine Reagent:

● Acetic Acid
● Concen. H2SO4
How to minimize interaction between DNA and
histones in experiment?
The experiment isolates the protein in an acidic environment, causing the histone
to precipitate.

Principle:
DNA sample is boiled under extremely acidic conditions which causes
depurination of the DNA followed by dehydration of deoxyribose sugar
 Diphenylamine Test
Results of groups 1-5

1% glucose Green

1% Ribose Light Blue

1% Deoxyribose Blue Violet

Crude DNA Violet

 DNA Extraction of White Onions

3 Stages:

1. Disruption of Cell membrane

2. Dissociation from and denaturation of proteins
3. Separation of DNA molecule from other cellular
components
 Sodium Dodecyl Sulfate

● Sodium salt of dodecyl hydrogen sulfate

● Amphiphilic
● 12 carbon tail (nonpolar) attached to sulfate group (polar)
● Clear Solution
 Sodium Dodecyl Sulfate

● Disrupts cell membrane

● SDS forms complexes with
lipids and proteins, causing
them to precipitate
● After adding onion, solution
is cloudy yellowish white
 Heating of 5% SDS and Blended Onion

● Heating ensures complete lysis of cells

● Inactivates DNAses
● Degrades proteins
● Suspends lipids
● Breaks down celluloid material
● Intracellular components are released
● No change in color
 Papain

● Enzyme that breaks up proteins

● When papain is added to the solution, it allows DNA to be separated from
proteins
● Solution turns slightly lighter yellowish white after adding grinded papain
tablet
 Ice Bath

● DNases are present in the cell’s cytoplasm

● Once the nuclear membrane is destroyed by homogenizing solution, DNA is
susceptible to DNases
● Cooling down the solution slows down process of degradation
● No change in color
 Filtration

● Separates cellular debris

● Small molecules flow through
● Solution is cloudy white
 Ice Cold Ethanol

● DNA is not soluble in ethanol

● Ice cold ethanol reduces enzyme activity and yields more DNA precipitate
● Clear white solution with white precipitate
 Spooling the DNA
● DNA comes as precipitate with mucus like texture when extracted
● DNA are polymers of nucleotides
● If DNA was powder like, we would not be able to see the DNA
References
National Human Genome Research Institute. (n.d.). Gene. Retrieved from:
https://www.genome.gov/genetics-glossary/Gene

National Human Genome Research Institute. (n.d.). Histone. Retrieved from:

https://www.genome.gov/genetics-glossary/histone

National Human Genome Research Institute. (n.d.). Nucleosomes. Retrieved

from: https://www.genome.gov/genetics-glossary/Nucleosome

National Human Genome Research Institute. (n.d.). Nucleotides. Retrieved from:

https://www.genome.gov/genetics-glossary/Nucleotide