Lecture Learning Activity 1.

Extract’urricular Biotalknology

https://www.semanticscholar.org/paper/Laboratory-Exercise-DNA-Extraction-Techniques-for-Hearn-Arblaster/cd8f896109d0bddd26bc7a61ac9452813483be13

INTRODUCTION

DNA extraction is of central importance to biotechnology. This concept is the gateway to

multiple applications and technologies, from basic research to routine diagnostic and
therapeutic decisions. Extraction and purification are also important in determining the
unique properties of DNA, such as size, shape, and function.

It is imperative that every BSA student obtain basic competency in DNA extraction
because “if you do not know the basic, you will be sick.” Each
molecule is microscopic but when extracted in bulk, it can be visible to the naked eye as
a clump of threadlike opaque slimy mucus-like material. If and when quality and quantity
of extracted DNA are less excellent, they could appear as hazy thin-like cream colored
stuff. If and when dirty and highly associated with proteins, they could appear as
brownish clumps (Jamago, 2021). In spite of resource limitations, every student should
be able to extract the DNA of his/her plant samples using basic materials available at
home.

Objectives

At the completion of this learning activity, the students are expected to:
1. demonstrate individually how to extract DNA using simple procedures,
equipment and materials;
2. and provide evidence of their extracted DNA from plant species and specimens
of their choice;
3. discuss the function of each material used; and
4. understand the basic structure of plant cells

MATERIALS AND METHODS

1. Every student shall decide on what plant species and plant part to use for DNA
extraction (e.g., mungbean seeds).
2. Prepare the following materials:
o 1 ripe table banana (e.g., latundan) and your choice of plant materials
(washed and clean: if leaves, make sure to use young leaves) – for
banana, precut into small pieces; for seeds, presoak in water overnight;
for leaves or flowers or stems, precut into small
pieces);
o salt (non-iodized, about 5 g),
o distilled water (bottled water will do, about 500 ml, preferably
chilled if possible),
o 70% ethyl alcohol (about 100 ml, preferably chilled* - put in the freezer the
night before use),
o clear shampoo or liquid detergent (about 10 ml per extraction
sample),
o beaker if available (if not, use a clear glass for drinking),
o resealable clear bag (like Ziploc) but if none, use a thicker clear plastic
cellophane that can be tightened
o strainer with fine meshing or if none, use a clean cheesecloth (or cotton)

o popsicle stick (wooden)

o water bath (boil tap water and put in a container & cover, its temperature
should be around 65°C = not too hot but not lukewarm either)
 o tape (masking tape or scotch tape) or use a rubber band
*if no refrigerator, soak in a container with ice

3. Prepare the extraction solution (this should be enough for several attempts or
trial and error but make sure you have extra banana and your other plant
tissues): mix 10 ml clear shampoo or detergent (if no graduated cylinder, 10 ml is
about 1 tablespoonful i.e. regular-sized spoon) + 1.5 g salt (a pinch) + 90 ml
distilled water. Stir very slowly to avoid foaming (or bubbling). Set aside in a
bottle or container.
4. Pour 20 ml of your extraction solution (or extraction buffer) into a medium size
zipper bag. Close the bag while squeezing out the air.

https://www.istockphoto.com/photos/ziplock-bag

5. Place the bag(s) into the hot water bath for about 10-15 minutes, making sure the
fruit solution is fully beneath the water line. Occasionally shake the bag to evenly
distribute the heat.
6. Move the bag (with mashed plant tissue) into another container with ice cold
water (ice bath) for 1 minute. Remove the bag and mix again the solution
carefully. Repeat about 5x.
7. Use your fine-meshed strainer or place your cheesecloth or cotton cloth over a
container. Tape the sides of the cloth to the container or use a rubber band to
secure the cloth. Filter the mixture through the cloth. Let the solution drain for 5
minutes.
 8. Using a tablespoon, transfer some of the filtered solution into another clean thin
glass. Slant the glass a little and add about 5 ml of chilled ethanol on the side of
the glass. Do not move or shake the glass.
9. Let the mixture sit for about 2-3 minutes without disturbing it. The DNA should
appear as a clump of transparent, slimy white mucus which can be gathered
using a popsicle stick and transferred to a small clear clean glass.
Congratulations!
10. Document your DNA extraction process (take photos or record a video).
11. Guide questions to answer:
o Why do you need to “mash” the plant tissue (or blend if you have a
blender)?
o Why is shampoo (or a liquid detergent) used? o
What is the purpose of using salt?
o Why is it necessary to use chilled and distilled water? o
Why is it necessary to cool down your mixture?
o What is the purpose of using ethanol?
o Why is it necessary to use cold or chilled ethanol?
o Why is it not advisable to use room temperature alcohol? o
Which plant specimen yielded more DNA?
o Describe your 2 DNA samples (banana and your chosen plant specimen)
or differentiate. Make sure to have photos of each.
12. When the work is done, always provide a simple discussion and conclusion in
your pre-recorded reports. If you have recommendations, you may also include
these.
13. To submit your pre-recorded reports, attach it to the assigned class work and
click TURN IN.
14. Filename of report should follow this suggested format: AGRIC45 LLA1
Report_Name of Student

REFERENCES:

✓ Jamago, JM. 2021. Up-close and personal with plant DNA. PB 201A (Advanced
Crop Genetics) Laboratory learning activity 1. 5p

✓ Online sources:
• https://drive.google.com/file/d/10S0nvg3yijyvhrvDHZuXUvbsbLqjT31A
/view
• https://www.youtube.com/watch?v=67KXatgoNKs
• https://whatisbiotechnology.org/index.php/science/summary/extract ion/dna-
extraction-isolates-dna-from-biological-material