PHINMA – RIZAL COLLEGE OF LAGUNA

School of Psychology

PLANT DNA EXTRACTION

I. Abstract
DNA extraction is an important step in many molecular biology research, such as
genetic trait analysis and organism identification. In this investigation, we looked into
the chemical extraction of DNA from plants as a preliminary study for future genetic
analysis. The goal of this study was to assess the yield and purity of DNA extracted from
different plant species, and to establish which approach was the most efficient and cost-
effective.
This lab report provides the results of a chemical approach for extracting DNA from
spinach and mango leaves. The purpose of this experiment was to evaluate the
efficiency and effectiveness of this extraction procedure, as well as to identify the
variety of plant that produces the highest DNA content. We examined the efficiency and
purity of DNA extraction from spinach and mango leaves, as well as the impact of
various doses of chemical reagents used in the extraction process. The findings of this
experiment revealed that the efficiency and purity of extracted DNA varied depending
on the kind of plant and extraction method. DNA outputs also varied, with spinach
leaves producing the highest concentration of DNA. Our findings highlighted the need of
selecting an effective extraction method for specific plant species, as well as carefully
evaluating the efficiency and purity of the extracted DNA in order to reduce DNA loss
throughout the extraction process.

II. Introduction
DNA extraction is a crucial procedure in molecular biology, which allows scientists to
separate and analyze the genetic material of organisms. It is a very important tool in a
number of scientific disciplines such as genetics, biotechnology, and plant breeding. The
knowledge of the process of DNA extraction from plant materials is necessary for the
investigation of plant genetics and the creation of new varieties with desirable
characteristics. In this laboratory experiment, we aimed to extract DNA from two
different plant species: Spinach leaves and Mango leaves. The aim was to compare the
efficiency and features of the DNA extraction process between these two plant sources.
Through analysis of the quantity of the extracted DNA and its quality, we can learn
about the efficiency of the extraction method and determine its feasibility for the use in
downstream molecular techniques.
The extraction procedure was rather complex, consisting of several steps such as
mechanical disruption of the plant cells, the addition of different reagents to release the
DNA, and the precipitation of the DNA using ethanol. This process was aimed at
obtaining visible and intact DNA samples that can be analyzed and used in future
experiments. The selection of Spinach and Mango leaves as plant sources for DNA
extraction was due to their availability and the fact that they may have different DNA
contents and characteristics. Spinach, which is renowned for its high nutritional value, is
likely to have a substantial amount of DNA. However, Mango leaves having different
morphology and composition might have different challenges and variations in the DNA
extraction process.
Through comparing the results of DNA extraction from Spinach and Mango leaves,
we can estimate the effectiveness of the extraction procedure and see if there are any
differences in the yield or quality between these two plant materials. This knowledge
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

will shape our knowledge of the factors that affect DNA extraction from various plant
species and inform future studies on plant genetics and biotechnology.

III. Procedure
This experimental approach demonstrates how to extract DNA from spinach and
mango leaves with a commercial kit. The goal of this experiment was to quantify the
yield and purity of DNA extracted from these two plant species, as well as to assess the
extraction method's efficacy. This knowledge is critical for molecular biologists and
other researchers that conduct DNA studies. The technique consists of multiple steps,
which will be discussed in full below.
1. We begin by weighing between 50 and 100 grams of the spinach leaves to start the
experiment then put it in a mortar pestle. After that, we pour around 100 ml of ice-cold
water into the pestle and half a teaspoon of common salt before we blend it all
together.
2. Using a funnel with a handkerchief, we pour the combined mixture into a beaker for
filtering.
3. To the filtrate in the beaker, we add a teaspoonful of dishwashing liquid soap, and
gently mix the mixture, then we leave the mixture for ten minutes to stand.
4. After ten minutes or so, we poured out 10 milliliters of the dishwashing liquid soap from
the beaker into a test tube.
5. After that, we prepare and pour in around 5 ml of pineapple juice. To combine the two
solutions, we gently shake the test tube and await the digestive reaction for ten
minutes.
6. Now, with the test tube tilted, we carefully add 5 ml of ethanol using a syringe into a
glass rod so that the alcohol softly runs down the inner wall of the tube and forms a
layer on top of the plant extracted that has been broken down.
7. We look at the interface where the two liquids meet. We notice a white gelatinous
strings or clumps which is the DNA after it precipitates in the alcohol.
8. We spool the resulting DNA onto the needle of the syringe to separate and remove it.
9. We made the same procedure for the mango leaves.

IV. Materials
This experimental procedure used the following materials for the extraction of
DNA from spinach and mango leaves. The materials used in the experiment were
both provided by the laboratory and the members who performed the experiment.

 Mango leaves
 Spinach leaves
 Cold water
 Salt
 Pineapple Juice
 Liquid Soap and Detergent Powder
 Tablespoon
 Teaspoon
 Handkerchief
 Funnel with filter paper cone
 Mortar and Pestle
 Weighing Scale
 250 ml beaker
 Test tube
 Test tube rack
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

 Glass rod
 5 ml or 10 ml syringe
 Ethyl Alcohol (100%)
 Stereoscopic microscope

V. Results
This experiment demonstrates how spinach and pineapple juices break down
DNA into its simplest cellular form. The salt helps the DNA stick together, and when
it reacts with alcohol, it becomes undissolved. This precipitation process pulls
additional strands, causing DNA to emerge. The experiment also demonstrates how
common salt removes proteins from the cell and DNA, making them more
hydrophobic. This method is widely used in molecular biology to separate and purify
DNA from biomolecules.
During the experiment, we assessed the efficiency and effectiveness of the DNA
extraction method for both spinach and mango leaves. The findings revealed that
DNA extraction from spinach leaves was more efficient and produced greater DNA
concentrations than DNA extraction from mango leaves. The differences in the yields
of DNA between the two plant types could be attributed to the differences in the
physiology of the leaves, such as their composition and structure. It is possible that
the higher concentration of contaminants and fiber in mango leaves compared to
spinach leaves could have inhibited the extraction process, leading to the lower
yields of DNA. The spinach leaves extraction is more visible under the microscope as
it shows a yellowish substance and a bit of bubbles on it from the dishwashing soap
that were used from the mixture. While the mango leaves weren’t apparent and just
shows a plain, white, almost like nothing under the microscope. This is most likely
because we weren’t able to extract the DNA completely. There were some issues
with extracting DNA from mango leaves. We had limited success extracting DNA
from mango leaves since the technique was time-consuming and required several
additional washing to remove contaminants. This resulted in a longer extraction
time, reducing the efficiency of the extraction procedure.
Overall, our findings demonstrated that the chemical approach utilized for DNA
extraction was effective for both spinach and mango leaves, but was more efficient
for spinach. The efficiency of DNA extraction from mango leaves, a mix of dried and
fresh leaves, was lower than that of spinach leaves. It is probable that the fiber
content of mango leaves contributed to the reduced extraction yield. Further testing
is required to enhance the mango leaf extraction procedure, particularly when fresh
leaves are used. The efficiency and purity of the extracted DNA are key variables in
downstream analysis, such as DNA sequencing, and the findings of this work will
help to create efficient DNA extraction methods for various plant species.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

VI. Discussion
It is challenging to extract plant DNA. Extracting DNA from animals is simpler.
Because plants have cell walls, they must be mechanically ground in a mortar and
pestle or in liquid nitrogen. Since animals lack cell walls, mechanical grinding is not
necessary.
Several factors can influence plant genomic DNA isolation efficiency. The age and
growth stage of the plant tissue are crucial factors to consider, as older or highly
mature tissues may have more complex cell walls, making DNA extraction more
challenging. It is preferable to use young, actively growing tissues for DNA isolation.
Furthermore, the concentration of DNA in plant cells might be low and vary
depending on the species and tissue type, therefore improving the extraction
procedure can improve the yield and purity of the DNA extracted. Another aspect is
cell viability; dead or dying cells can release enzymes that destroy DNA, so working
with live cells is critical for efficient DNA extraction. The effectiveness of DNA
extraction can also be affected by the kind of cell rupture used, such as grinding or
chemical treatments, because plant cell walls can be a considerable obstacle to
extraction. Finally, the choice of DNA purification procedures, such as
chromatography, precipitation, and column-based purification, might influence the
purity and quality of the extracted DNA. To improve the efficiency of plant genomic
DNA isolation, consider the factors mentioned above and optimize the extraction
method accordingly, such as selecting young, actively growing tissue, optimizing cell
rupture methods, adjusting DNA purification methods, controlling temperature and
pH, and using plant-specific buffers.
DNA extraction is a critical step in many types of biological study, including
molecular biology, genetic engineering, plant breeding, and ecology. The capacity to
extract DNA from plants enables researchers to sequence and analyze plant
populations' genetic material, track changes in the plant genome over time, and
create new plant kinds with desirable features. DNA extraction is significant in plant
research because it allows access to the genetic information stored in the plant's
genome. This genetic information can be used to discover the genes that cause
desired features like greater drought tolerance or insect resistance, as well as to
monitor the spread of genetically modified organisms in the environment. In
addition to its research applications, DNA extraction from plants is essential in the
biotechnology and agriculture industries. DNA extraction, for example, can be used
to produce molecular diagnostic tools for identifying plant diseases or to establish
novel plant kinds through genetic engineering. Overall, DNA extraction from plants is
a fundamental technique that offers a foundation for many disciplines of biological
research. It is essential for expanding our understanding of the plant genome and
generating novel applications in biotechnology, agriculture, and other sectors.
When it comes to extracting chemicals from spinach leaves, selecting the
appropriate solvent is critical. The solvent used is determined by the individual
substances to be extracted, as well as the solvent's characteristics. Acetone,
methanol, ethanol, and water are some of the most often utilized solvents for
spinach leaf extraction. Acetone is a highly effective solvent for extracting both polar
and non-polar chemicals from spinach leaves. The solvent's capacity to dissolve both
polar and non-polar molecules makes it an adaptable alternative for extracting a
wide spectrum of chemicals from leaves. Furthermore, acetone is affordable and
simple to use, making it a popular choice for laboratory studies and commercial
uses. Methanol is another good solvent for extracting polar components from
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

spinach leaves, like vitamins and antioxidants. It has a lower volatility than acetone
and can be used to extract heat-sensitive or acetone-degradable chemicals.
However, methanol should be handled with caution due to its flammability and
toxicity. Ethanol is also a popular solvent for extracting chemicals from spinach
leaves since it can dissolve both polar and non-polar molecules. Ethanol is less
volatile than methanol, making it safer to handle, and it is commonly utilized in the
food and beverage sectors. Water is another method for removing chemicals from
spinach leaves. While it may not be as effective as other solvents at dissolving some
molecules, it is an environmentally benign method for extracting hydrophobic
substances from leaf tissue. Water can also be used to remove chemicals that might
otherwise be damaged by more strong solvents.
The extraction period has a substantial impact on the yield of extracted
components from spinach leaves because it influences the release of compounds
from the plant tissue. In general, increasing the extraction period can result in higher
yields of extracted chemicals since it allows the solvent to release the compounds
from the leaf tissue. However, the ideal extraction period will be determined by the
solvent employed and the leaf tissue's properties. If the solvent is too aggressive or
the extraction time is too long, the extracted components will degrade. However, if
the solvent is too weak or the extraction time is too short, all of the chemicals in the
plant tissue may not be released efficiently. In general, leaves with thick, fibrous cell
walls require longer extraction times since they are more difficult to penetrate.
Softer and fleshier leaves, such as spinach, may require shorter extraction durations
to avoid excessive degradation.
The ideal temperature for extracting chemicals from spinach leaves varies based
on the solvent employed and the specific compounds being extracted. However, the
extraction procedure is normally carried out at ambient temperature, which is
approximately 25°C (77°F). Many of the chemicals found in spinach leaves are stable
at room temperature and can be removed without undergoing significant
degradation. Furthermore, room temperature is a safe and easy to control
temperature for the extraction procedure. However, some chemicals may require
greater or lower temperatures to extract successfully. Certain vitamins, such as
Vitamin C, deteriorate at higher temperatures, therefore room temperature is
frequently the best option for extracting them. On the other hand, some compounds
may be more soluble at higher temperatures, hence raising the temperature during
the extraction process may enhance their yield. In general, the best temperature for
extracting chemicals from spinach leaves should be found by experimentation,
taking into account the individual compounds being extracted as well as the
properties of the solvent utilized.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

VII. Conclusion
The experiment successfully demonstrated the breakdown of DNA into its
simplest cellular form for spinach leaves using pineapple juices, aided by salt and
alcohol precipitation. The method effectively separated and purified DNA from
biomolecules, showcasing its utility in molecular biology. While the efficiency of DNA
extraction was higher for spinach leaves compared to mango leaves, likely due to
differences in leaf physiology, challenges were encountered with mango leaves
extraction, attributed to its higher fiber content. Further optimization is needed for
mango leaves extraction, especially with fresh leaves, to improve efficiency. The
findings underscore the importance of efficient DNA extraction methods for various
plant species, crucial for downstream analyses like DNA sequencing. This study
contributes valuable insights towards developing effective extraction techniques for
diverse plant DNA studies.
The process of extracting DNA from plants presents unique challenges compared
to animals due to the presence of cell walls, necessitating mechanical grinding for
access. Factors such as tissue age, cell viability, and DNA concentration influence
extraction efficiency. Selecting appropriate methods, such as mechanical or chemical
cell rupture, and purification techniques is crucial for optimal results. DNA extraction
from plants plays a pivotal role in various fields, including molecular biology, genetic
engineering, and ecology, enabling research on genetic traits and biotechnological
applications.
Extraction duration and temperature impact yield and degradation, with longer
durations aiding compound release but risking degradation and optimal
temperatures varying based on the solvent and compounds being extracted. Overall,
when we put the DNA of spinach in the microscope, it was successful, while the
mango leaves didn't see much because there was so little DNA that the microscope
saw in the mango leaves, careful consideration of these factors is essential for
successful chemical extraction from spinach leaves, enabling a broad range of
applications in research and industry.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

VIII. References
Spencer, Donald and Whitfeld, Paul. (2004) The characteristics of spinach chloroplast
DNA polymerase. Archives of Biochemistry and Biophysics;
https://www.sciencedirect.com/science/article/abs/pii/0003986169903920

Krause, Kirsten and Krupinska, Karin. (2001) Molecular and functional properties of highly
purified transcriptionally active chromosomes from spinach chloroplasts. Physiologia
Plantarum; https://onlinelibrary.wiley.com/doi/abs/10.1034/j.1399-3054.2000.100211.x

Scobeyeva, V. and Omelchenko, D. (2018)

Comparison of Some Plant DNA Extraction Methods. Russian Journal of Genetics;
https://link.springer.com/article/10.1134/S1022795418050095

Rogstad, Steven and Keane, Brian. (2001) DNA extraction from plants: The use of
pectinase. Plant Molecular Biology Reporter;
https://link.springer.com/article/10.1007/BF02772833

Huang, Zangbao and Ren, Hui. (2020) A modified method of Total RNA Isolation for
Mango Leaf Tissues. An International Journal for Bioscience Methods and Technologies;
https://bioscipublisher.com/index.php/bm/article/view/3744

Maaty, Nadia and Oraby, Hanaa. (2019) Extraction of high-quality genomic DNA from
different plant orders applying a modified CTAB-based method. Bulletin of the National
Research Centre; https://bnrc.springeropen.com/articles/10.1186/s42269-019-0066-1

Jaime, Laura and Vasquez, Erika. (2014) Extraction of functional ingredients from spinach
(Spinacia oleracea L.) using liquid solvent and supercritical CO2 extraction. Journal of the
Science of Food and Agriculture; https://onlinelibrary.wiley.com/doi/10.1002/jsfa.6788

Derrien, Maelle and Badr, Ashraf. (2017) Optimization of a green process for the
extraction of lutein and chlorophyll from spinach by-products using response surface
methodology (RSM). Food Science and Technology;
https://www.sciencedirect.com/science/article/abs/pii/S0023643817300105

Azmir, J. and Zaidul, I. (2013) Techniques for extraction of bioactive compounds from
plant materials: A review. Journal of Food Engineering;
https://www.sciencedirect.com/science/article/abs/pii/S0260877413000277#:~:text=In%
20hydrodistillation%2C%20first%2C%20the%20plant,bioactive%20compounds%20of%20
plant%20tissue.

Leite, Ana and Ferreira, Ana. (2018) Cloud point extraction of chlorophylls from spinach
leaves using aqueous solutions of non-ionic surfactants. National Library of Medicine;
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6161820/
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

IX. Documentation

To start off the experiment, the students weigh the spinach leaves between50 and 100
grams.

After that, we pour around 100 ml of ice-cold water into the pestle and half a teaspoon of common
salt before we blend it all together.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

Afterwards, the spinach leaves were After achieving the gelatinous texture,
grinded and crushed using the pestle the students added a half teaspoon of
until they achieved a gelatinous texture. common salt and blended it.

By using a funnel with a handkerchief, the students poured the combined mixture into the
test tube for filtering and transferred it into a beaker.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

After filtering the mixture, the students After 10 minutes, the students poured 10
poured a spoonful of dishwashing soap into ml of the mixture from the beaker into the
the mixture and gently mixed the mixture. test tube.
The students waited for 10 minutes for the
next procedure.

After transferring the mixture to the test While the test tube was tilted, the students
tube, the students poured in around 5 ml of carefully poured 5 ml of ethyl alcohol using
pineapple juice. In order to combine the a syringe.
two solutions, the student used a stirrer
rod. Then, they waited for 10 minutes for
the digestive reaction.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

The DNA started to precipitate in the The DNA obtained from spinach leaves
alcohol as gelatinous clumps. was taken using a needle and transferred
to the glass slide.

The extracted DNA was flattened This is what the DNA extracted from
using the needle and put under the spinach looks like under the microscope.
microscope.

On to the next part: The students weigh Beforehand, the students decided to
the mango leaves between 50 and 100 crush off the mango leaves to make the
grams. next procedure easier.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

After weighing, the students put the mango leaves in a pestle or mortar and poured 100
ml of cold water over them.

Afterwards, the mango leaves By using a funnel with a

were grinded and crushed using handkerchief, the students poured
the pestle. Then, the students the combined mixture into the
added a half teaspoon of common test tube for filtering.
salt and blended it.

After filtering the mixture, the students poured a

spoonful of dishwashing soap into the mixture
and gently mixed the mixture. The students
waited for 10 minutes for the next procedure.
 PHINMA – RIZAL COLLEGE OF LAGUNA
School of Psychology

After 10 minutes, the students After transferring the mixture to

poured 10 ml of the mixture from the test tube, the students poured
the beaker into the test tube. in around 5 ml of pineapple juice.
In order to combine the two
solutions, the student used a
stirrer rod.

The students waited for 10 minutes While the test tube was tilted, the
for the digestive reaction. students carefully poured 5 ml of ethyl
alcohol using a syringe.

The students waited for the DNA to

precipitate.