GUÍA DE LABORATORIO

Fecha de
DEPARTAMENTO DE CIENCIAS NATURALES Y Código:
emisión:
EDUCACIÓN AMBIENTAL GCU-LAB-03
9-Jun.-10
GIMNASIO LOS PINOS
Elaboro: Jefe Ciencias Aprobó: Dir. SGC Versión 1 Página 1 de 3

LABORATORY GUIDE # 2

VEGETABLE AND ANIMAL DNA EXTRACTION

III TERM – BIOLOGY

GRADE 8

OBJECTIVE

1. Recognize DNA as life molecule in living things.

FRAMEWORK1

What is DNA Extraction?

Simply put, DNA Extraction is the removal of deoxyribonucleic acid (DNA) from the cells or viruses
in which it normally resides.

What is it used for?

Extraction of DNA is often an early step in many diagnostic processes used to detect bacteria and
viruses in the environment as well as diagnosing disease and genetic disorders. These techniques
include but are not limited to -
 Fluorescence In Situ Hybridization (FISH): FISH is a molecular technique that is used,
among other things, to identify and enumerate specific bacterial groups.
 Terminal Restriction Fragment Length Polymorphism (T-RFLP): T-RFLP is used to identify,
characterize, and quantify spatial and temporal patterns in marine bacterioplankton
communities.

 Sequencing: Portions of, or whole genomes may be sequenced as well as extra

chromosomal elements for comparison with existing sequence in the public data base.

How does it work?

(Outline of a basic DNA Extraction - )

1. Break open (lyse) the cells or virus containing the DNA of interest-
This is often done by sonicating or bead beating the sample. Vortexing with phenol
(sometimes heated) is often effective for breaking down protienacious cellular walls or
viral capsids. The addition of a detergent such as SDS is often necessary to remove lipid
membranes.
 GUÍA DE LABORATORIO
Fecha de
DEPARTAMENTO DE CIENCIAS NATURALES Y Código:
emisión:
EDUCACIÓN AMBIENTAL GCU-LAB-03
9-Jun.-10
GIMNASIO LOS PINOS
Elaboro: Jefe Ciencias Aprobó: Dir. SGC Versión 1 Página 2 de 3

2. DNA associated proteins, as well as other cellular proteins, may be degraded with the
addition of a protease. Precipitation of the protein is aided by the addition of a salt such as
ammonium or sodium acetate. When the sample is vortexed with phenol-chloroform and
centrifuged the proteins will remain in the organic phase and can be drawn off carefully.
The DNA will be found at the interface between the two phases.

3. DNA is the precipitated by mixing with cold ethanol or isopropanol and then centrifuging.
The DNA is insoluble in the alcohol and will come out of solution, and the alcohol serves as
a wash to remove the salt previously added.

4. Wash the resultant DNA pellet with cold alcohol again and centrifuge for retrieval of the
pellet.

5. After pouring the alcohol off the pellet and drying, the DNA can be re-suspended in a
buffer such as Tris or TE.

6. Presence of DNA can be confirmed by electrophoresing on an agarose gel containing

ethidium bromide, or another fluorescent dye that reacts with the DNA, and checking
under UV light.

MATERIALS

*NOTE: BRING PER LAB GROUP, THE HIGHLIGHTED MATERIALS. WITHOUT THESE MATERIALS YOU
WILL NOT BE ALLOWED TO ENTRANCE.

ORGANIC Wire gauze (malla de asbesto)

Vegetable or plant
REACTIVES
GLASS AND OTHERS Soap
Mortar and pestle Salt (NaCl2)
2 Beakers Isopropyl alcohol
6 Plastic spoon Lysis solution
6 plastic cup Natural Papaya juice (without sugar).
Filter paper
Test tube SECURITY
10 pasteur pipette or eyedropper Lab robe
Test tube holder Latex or nitrile gloves
Buccal swab Lab mask
Microcentrifuge

METHODOLOGY

ANIMAL DNA EXTRACTION2:

1. Watch the virtual lab on the web site: http://learn.genetics.utah.edu/content/labs/extraction/

and based on it do the fluid diagram.
 GUÍA DE LABORATORIO
Fecha de
DEPARTAMENTO DE CIENCIAS NATURALES Y Código:
emisión:
EDUCACIÓN AMBIENTAL GCU-LAB-03
9-Jun.-10
GIMNASIO LOS PINOS
Elaboro: Jefe Ciencias Aprobó: Dir. SGC Versión 1 Página 3 de 3

VEGETABLE DNA EXTRACTION3:

1. Put 1/2 cup of distilled water and the vegetable into the mortar and pestle. Crush it, making
sure the vegetable is completely pulverized. Pour the mixture into a beaker.
2. Mix 1 teaspoon of soap with 1/4 teaspoon of salt in a plastic cup. Add 2 tablespoons of distilled
water.
Stir gently to avoid creating a foam. Continue for a few minutes until the soap and salt are
dissolved.
3. Add 2 tablespoons of the vegetable mixture to the cup containing the soap solution. Use a
spoon to stir the mixture for at least 10 minutes.
4. Insert a filter into a clean plastic cup so it does not touch the bottom of the cup. If necessary,
tape the sides of the filter to the cup.
5. Pour the mixture from step 3 into the filter. After 10 minutes, some liquid, called the filtrate,
should have collected in the bottom of the cup. Gently stir the mixture in the filter and let it sit for
another minute. Remove the filter and set it aside.
6. Get a test tube of cold alcohol. Use a pipette or eyedropper to collect your filtrate. Add it to the
alcohol.
7. Place the test tube with the alcohol and filtrate in a beaker or test tube holder. Let it sit
undisturbed for about four minutes. Do not shake. The white material coming out of solution as a
precipitate is DNA.
8. Dip the glass rod into the tube, slowly rotating it to spool out the vegetable’s DNA.

BIBLIOGRAPHY

1. George Rice, Montana State University (2012). DNA Extraction. Retrieved July 31,
2012, from
http://serc.carleton.edu/microbelife/research_methods/genomics/dnaext.html

2. Genetic Science Learning Center (1969, December 31) DNA Extraction Virtual Lab.
Learn.Genetics. Retrieved July 31, 2012, from
http://learn.genetics.utah.edu/content/labs/extraction/.

3. Extracting DNA from banabas. ©2005 WGBH Educational Foundation. NOVA and NOVA
science NOW are trademarks of the WGBH Educational Foundation. Retrieved July 31, 2012,
from http://www.pbs.org/wgbh/nova/teachers/activities/pdf/3214_01_nsn_01.pdf