PEDROSA, JOANRD T.

BSED BIOSCI 4

VIRTUAL CHEMISTRY LAB: DNA EXTRACTION

Open the attached swf file. Note: You’ll be needing an access to a laptop or desktop. This won’t
need internet connection since this is a shockwave player file. But if you want to access it
through the internet, visit this link: https://learn.genetics.utah.edu/content/labs/extraction/

PROCEDURES:
1. As you open the simulator, click “start lab”.
2. Read the instructions on what to do in each slide. Use the “next” and “back” button to
get from slide to slide. In this experiment you will collect cheek cells from a person in
order to isolate (separate) DNA from the cheek cells. Follow along and answer these
questions:
a. Why do scientists isolate DNA? List some reasons:
-Genetic testing
-Body identification
-Analysis of forensic evidence

b. What is DNA Extraction?

-DNA Extraction is typically the first step in a longer laboratory process. This is
an important part of that process because the DNA first needs to be purified away
from proteins and other cellular contaminants. We need cells because that’s where
the DNA is inside almost every cell in our bodies is a nucleus, and inside each
nucleus is about two meters of DNA.

3. Watch carefully what happens in slide #4. Explain what happens to slide #4 below:

At the center of the cell, nucleus is located. Inside the nucleus, where is what we called
chromosomes.chromosomes are made up of histones where DNA is located.
4. Continue the simulation. Answer the following:
c. What are the equipment to be used in the virtual experiment?
- Warm water bath, centrifuge, buccal swab, micro pipettors, sample tubes,
lysis solution, concentrated salt solution, resuspension buffer, ethanol,
isopropyl alcohol

d. What are the steps in the DNA extraction done in this virtual lab?
- 1. Collect cheek cells
- 2. Burst cells open to release DNA
- 3. Separate DNA proteins and debris
- 4. Isolate concentrated DNA
e. What does “Lysis solution” do?
- The term "cell lysis solution" is sometimes, though not always, used
interchangeably with "lysis buffer." So it is useful to know the specific
ingredients of a chemical cocktail designed specifically to break down the cell
membrane without compromising the integrity of the cell contents.

f. What does the detergent do?

- The detergent disrupts the cell membrane and nuclear envelope, causing the
cells to burst open and release their DNA.

g. What is the protein called DNA is wrapped around?

- Histones, and the proteinase K outs apart the histones to free the DNA.

h. “The salt causes the ___proteins_____ and other cellular debris to

___clump______ together. After being spun in the micro centrifuge, the
_DNA______ remains distributed through the liquid.

i. Because the DNA is not __soluble _____ in isopropyl alcohol, if comes out of the
solution. You can see __the____ clumped with your naked eye!

5. Continue the activity by answering the following guide questions. Use your schema on
other concept relating to qualitative tests.

a. Differentiate RNA from DNA using the table below:

Criterion DNA RNA
Based on their function Hold genetic information
Based on their sugar composition Deoxyribose
Based on their pyrimidine base Cytosine
composition
Thymine
Based on their structure Deoxyribonucleic acid
BASE PAIRING Double stranded sugar phosphate
Thymine, cytosine, adenine, guanine
b. Indicate the possible results of the nucleic acid in the following qualitative tests.
Note the result as positive or negative to the test and provide justification.
Qualitative tests DNA
Result: The presence of smaller
carbohydrates like glucose can be
Benedict’s Test determined by
using Benedict's solution. Benedict's solution
has copper ions (Cu2+) that have a light blue
color. ... The presence of DNA will turn a
clear solution blue. The more DNA present
the darker the color.
Justification: Due to its deoxyribose sugar,
which contains one less oxygen-containing
hydroxyl group, DNA is a more stable
molecule than RNA, which is useful for a
molecule which has the task of keeping
genetic information safe.
Result: The chemical reaction causes the
Biuret solution to turn from a light blue to
Biuret’s Test purple if proteins are present. ... The
presence of DNA will turn a clear solution
blue. The more DNA present the darker the
color. Another nucleic acid, RNA, will turn
green.
Justification: NaCl, ethanol and detergent
Transcribes and regulates the
genetic information
Ribose
Cytosine
Uracil
Ribonucleic acid
Usually single stranded sugar
phosphate
Uracil, cytosine, adenine, guanine
RNA
Result: Benedict's solution has
copper ions (Cu2+) that have a
light blue color. ... The
presence of DNA will turn a
clear solution blue. The more
DNA present the darker the
color. Another nucleic acid,
RNA, will turn green.
Justification: RNA, containing
a ribose sugar, is more reactive
than DNA and is not stable in
alkaline conditions. RNA’s
larger helical grooves mean it
is more easily subject to attack
by enzymes.
Result: Proteins can be
detected by their reactions
with Biuret's reagent. ... RNA
contains ribose sugars and
does not react with the Dische
diphenylamine reagent. The
intensity of the blue color is
proportional to the
concentration of DNA.
Justification: NaOH and
C2H5OH (ethanol) with conc.
 HCI

Result: brown sol’n Result: Yellow precipitation

Ammonium Molybdate Justification: hosphate group is present in Justification: commercial
Test both RNA and DNA ,during experiment RNA gave positive test due to
phosphate ions reacts with ammonium presence of phosphate in
molybdate and in acidic medium it forms them.Purine bases test
blue colour complx.Lab DNA,commercial indicates presence of purines.
DNA,Na3PO4

Result: Brown sol’n Result: Gelatinous white

precipitation
10%NH4OH & Justification: The base units in DNA are
adenine, guanine, cytosine and thymine. ...
10%AgNO3 Test The hydrolysis products of DNA are
Justification: The hydrolysis
phosphoric acid, deoxyribose, the bases A,
products of RNA are
G, C and T. 6.
phosphoric acid, ribose and
bases A.G, C and U.

Reference:
Genetic Science Learning Center, The University of Utah (2006). Virtual Chemistry Lab: DNA
Extraction. Retrieved from: https://learn.genetics.utah.edu/content/labs/extraction/