PRACTICAL REPORT

Practical 4: DNA Extraction

Group Members :

Name Matrics Number

NURUL ALIA SHAFYKA BINTI REDZOAN A188275
NUR AYESHAH AMEERA BINTI NORAZILEE A188766
ARYSSA BINTI AZRI A189578
SHRRUTI A/P SURESH KUMAR A189624
 PRACTICAL REPORT
Practical 4: DNA Extraction

Group Members :

Name Matrics Number

NURUL ALIA SHAFYKA BINTI REDZOAN A188275
NUR AYESHAH AMEERA BINTI NORAZILEE A188766
ARYSSA BINTI AZRI A189578
SHRRUTI A/P SURESH KUMAR A189624
 Date : 10/11/2021
Lecturer : PROF. MADYA DR. GOON JO AAN​Results
1. Write down the results of this experiment in the table below. Discuss the
significance of A260/A280 ratio.

Absorbance Result significance:indication of purity.

ratio of 1.7-2.0 ->
purtDNA
A260 3.026 3.981 ~2.0
pure RNA
->

A280 1.610 2. 145

A90:
ofDNA8
*

A230
Indary measure

perity
A260/ A280 1.880 1.86 RNA

2. Calculate the concentration of DNA in your blood sample.

Concentration of DNA in Sample = (50ng/µl) A260

= (50ng/µl)3.026
= 151.30 ng/µl

CanC A260*
DNA= 50
ng/mL
3.981 50ng/mL
=

=199ng/mL
Questions:

1. Explain the principle of DNA extraction.

DNA extraction is a method of extracting DNA from biological samples such as body
-

fluids, tissues and microorganisms. The purpose of DNA extraction is mainly for
diagnosis, prevention and treatment of diseases. The steps of DNA extraction
-

include cell lysis, protein precipitation, DNA precipitation and DNA hydration.
-
- -

Yo
DNA extraction protocols comprise of the basic steps of disruption of the cell
-

wall, cell membrane and nuclear membrane to release the DNA into solution
-

followed by precipitation of DNA while ensuring removal of the contaminating

-

biomolecules such as the proteins, polysaccharides, lipids, phenols and other

secondary metabolites.
Raising to room temperature adds heat that can provide a heating condition
-

which can help to denature proteins, eliminate nucleases, and cause the DNA
-
30
extracted from the cell, so that the DNA can serve as a template for the later
↳
-

amplification reaction. When protein is denatured and being degraded, it can easily
-

precipitate.
-

The cells in a sample are separated from each other physically during vortexing,
-

and put into a solution containing salt. The positively charged substrates in the salt
-

help protect the negatively charged phosphate groups that run along the backbone
-
-

of the DNA. DNA is released as these membranes are disrupted.

-

Then, protein precipitation solution is then added to separate DNA and other
-
-

cellular debris. Essentially, this is a high-salt buffer that lowers the solubility of
- -

proteins.
-

DNA is soluble in water but insoluble in the presence of alcohol, therefore when
e - e

alcohol is added, we can see the white thread which is the DNA.
-

Adding ethanol helps to wash the DNA pellet.

- -

Hydration the solution means adding DNA Hydration Solution to make the sample
become a solution from the pellet. So, it will be more accurate when we measure the
- >

absorbance using spectrophotometer and it helps to avoid inaccurate DNA

-

quantification
2. What are the major contaminants in DNA extracts? Describe the purity of your
sample.
Major contaminants in DNA extracts include proteins, polysaccharides, lipids, y from extraction
aut
phenols. The purity of our sample is at 1.880 which is in the optimal range of ~1.7- reay
1.9.
Fab

3. What is the purpose of

a. raising the sample temperature to room temperature before adding Protein
Precipitation Solution?
Increasing the temperature can increase the efficiency and enhance the yield. It can
also prevent the co-precipitation of salts which can occur at low temperatures.
Ifom
 W

b. hydrating the sample completely?

To ensure no DNA left out from the sample solution and dissolve back into the
solution to get an accurate spectrophotometry results which indicates a more
accurate DNA quantification.

minciple
DNAextraction
samples.
-.....

Its
purpose is for dx, prevention, I of
dis

cyss,
* protein membprecipitation, hydration ......

First
step is the disruption of cell it release DNAinto static
to

followed
by DNAprecipitation mering
removal

Eamirants,
rocurtemp provide and to denature proton.
raising to

dinucleases and extractDNAfrom the all so it

as
·
template for amplication. When protein is

denatured, easing precipitate.

itcan
vortexing.
alls in
sample separated physically though
as

DNA is released these membraces

of
disrupted. are

Proteinprecipitate is added
separate DNAand other
to

allular debris.

Anding alcouel help precipitate

pro to presence
leading
in
alcald.
of white thead as DNA is insoluble
adding hydration santo helps sample become scutic
from pellet. So the absorbance can be measured and
port
acid inaccurate
↓ quantificatio