8

Research 2 Activity
Sheet
Quarter 3 – MELC 5
Week 5
Laboratory Techniques and
Methods

REGION VI – WESTERN VISAYAS

Research 2
Activity Sheet No. 4- Laboratory Techniques and Methods
First Edition, 2021

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Introductory Message
Welcome to Research II!

The Learning Activity Sheet is a product of the collaborative

efforts of the Schools Division of San Carlos City and DepEd Regional
Office VI - Western Visayas through the Curriculum and Learning
Management Division (CLMD). This is developed to guide the learning
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learners meet the standards set by the K to 12 Basic Education
Curriculum.

The Learning Activity Sheet is self-directed instructional materials

aimed to guide the learners in accomplishing activities at their own pace
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also assist the learners in acquiring lifelong learning skills, knowledge, and
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For learning facilitator:

The Research 2 Activity Sheet will help you facilitate the leaching-
learning activities specified in each Most Essential Learning Competency (MELC)
with minimal or no face-to-face encounter between you and the learner. This will
be made available to the learners with the references/links to ease independent
learning.

For the learner:

The Research 2 Activity Sheet is developed to help you continue

learning even if you are not in school. This learning material provides you with
meaningful and engaging activities for independent learning. Being an active
learner, carefully read and understand the instructions then perform the activities
and answer the assessments. This will be returned to your facilitator on the
agreed schedule.
Name of Learner:
________________________________________________________
Grade and Section: ___________________________
Date:_____________________
 RESEARCH 2 ACTIVITY SHEET NO. 4
Laboratory Techniques and Methods

I. Learning Competency with Code

o Identify the standard methods and techniques used in performing

experiments (SSP_RS8-SA-III-o-q17).

II. Background Information for Learners

In making scientific research, designing and conducting an experiment is of
great importance. An experiment is a set of procedures that are used by the
investigator to be able to collect the necessary information that is needed to
verify or test the truthfulness of the hypothesis or to answer the research
questions. In an experiment, all the essential conditions are controlled so that the
results can only be attributed to the independent variable (Lahorra, 2020).

Experiments are done under controlled conditions so that the results can be
achieved. Important concepts must be considered in learning about designing
and conducting experiments.

Study the table below.

Set-up of the Experiment
Concepts Experimental Control + Control -
Controlled variables ▪ Kind of tomato ▪ Kind of tomato ▪ Kind of tomato
are factors that are plant plant plant
maintained in the ▪ Type of soil ▪ Type of soil ▪ Type of soil
experiment. This is ▪ Amount of ▪ Amount of ▪ Amount of
done to avoid other water water water
factors intervening
▪ Amount of ▪ Amount of ▪ Amount of
in the results.
sunlight sunlight sunlight
A dependent ▪ Growth of ▪ Growth of ▪ Growth of
variable is a factor pechay plants pechay plants pechay plants
or condition that in terms of in terms of in terms of
might be altered. height (cm) height (cm) height (cm)
The effect could be
observed and
measured.

An independent ▪ Organic ▪ Commercial ▪ Without

variable is a factor or fertilizer fertilizer fertilizer
condition that might
be affected. The
effect could be
observed and
measured.
Experimental unit If a researcher wants to find out the growth rate of
refers to the subject pechay plants after three weeks wherein they will
or object which is receive different treatments (with organic fertilizer and
introduced with a without fertilizer). Therefore, the pechay plants are the
stimulus and is then experimental unit.
measured.
Treatment is the Using the same example on the experimental unit, the
stimulus introduced treatments introduced are – (a) adding organic fertilizer
to the experimental on the soil and (b) without adding fertilizer on the soil.
subjects.
Repeated Take for example if researchers wanted to test a
measurements certain medication on patients with Diabetes II, they will
are the be first tested to measure their glucose level before
measurements they are given a specific treatment. After the treatment
that are made on has been implemented, patients will be tested again
each subject over and they might undergo nutritional counseling as well.
several periods in
some
experiments.
Repeated Using the same example above, the measurement
measurement design to be used is a crossover design – “two-period,
two-treatment” which means the research participants
design refers to the
are randomly assigned to receive two different
design which treatments: either A then B, or B then A.
includes repeated
measurements on
research where a
single treatment is Then also applicable to research studies that may
administered to a involve multiple factors affecting the results gathered
subject or it involves from it.
designs of treatment
that are modified
before each
measurement.
The purpose of the control group or control treatment is for comparison. The
results from the experimental treatment will be compared to the results of the
control group. This helps in making valid conclusions about how organic fertilizer
showed its effects on the growth of the plants. There are two types of control
groups in experiments: negative and positive control groups. A negative
control group receives a negative control treatment (this is without treatment
or a treatment that will not give an effect). A positive control group receives a
positive control
treatment, a treatment that is already known for a particular effect.

LABORATORY TECHNIQUES AND METHODS

Aside from doing the Scientific Method as a general method of conducting

any Science-related activities, laboratory methods and techniques are essentially
introduced since experiments are mostly done in laboratories. Cambridge
dictionary defines a laboratory as “a room or building with scientific
equipment for doing scientific tests for teaching Science, or a place where
chemicals or medicines are produced.” On the other hand, laboratory
techniques are procedures and practices that are required in utilizing various
equipment inside the laboratory.
Here are some of the laboratory techniques and methods that you need to
learn as you conduct your research study:

A. Microbiological Techniques
In microbiology, microorganisms like archaea, bacteria, fungi, protozoa,
viruses, etc. are being studied.
 The common microbiological techniques are discussed in detail below:

1) Sterilization
Microbiological media, a medium for growing bacteria or any other
microorganisms, must be sterilized by heating it in an autoclave (like a pressure
cooker) at 121°C for 15 minutes. This will kill all living organisms, i.e., spores.

2) Aseptic Techniques
This is done to lessen the chance of bacterial contamination. Furthermore,
this also involves disinfecting working area 4exposed to microbiological media
to minimize the possible contact by bacteria from the air and use flames to kill
bacteria that might enter the vessels after opening them.

3) Inoculation
This is the process by which bacteria are introduced to a media in various
means. For example (bacteria), from a drop in a heat-sterilized loop are
dispersed on the surface of the agar. This is the same with broth cultures. For
bacteria in a liquid, a sterile pipette is introduced to a Petri dish before the agar
is poured on the top (“pour plates”).

4) Incubation

This is the process of maintaining favorable conditions for growth and

development, i.e., microbiological culture.

The Petri dishes that contain the agar or tubes contain broth are incubated,
for example, a special apparatus is placed in a room with a fixed temperature
(usually at 37 °C, for pathogens, while for bacteria is 35 °C from the
environment). In school, incubation temperatures are set lower to minimize the
growth of potential pathogens.

B. Chemical

1) Spectrophotometry
This is a method by which you measure the amount of absorption and
transmittance of light. This is used widely for qualitative analysis in various
fields of Sciences (e.g., biochemistry, biology, chemistry, material and chemical
engineering, clinical applications, industrial applications, etc. ). A
spectrophotometer is a device that measures the number of photons (the
intensity of light) passing through a sample solution. This consists of two
devices: a spectrometer and a photometer. A spectrometer is a device that
disperses and measures light. On the other hand, a photometer consists of a
photoelectric detector that measures the intensity of light.

2) Extraction

This is the conversion of compound/s from a solid or liquid state into

another phase or state.
 Preparation of Crude Extracts from Plants

Studies about the medicinal properties of plants require the extraction of

crude compounds from plant parts such as leaves, flowers, and roots.
Knowledge on how to do the extraction process is necessary.

Step 1: Air Drying of Samples. After collecting the adequate amount of plant
5
samples, wash with clean water and dry at room temperature. The efficiency
of extracting crude compounds can be achieved when the plant samples are
airdried.

Step 2: Grinding the Dried Samples into Powder. The purpose of this step is
to turn samples into small pieces (powder) using a Wiley Mill or if you do not
have one, you can use a pair of scissors. Compounds can easily dissolve in the
solvent when the plant sample is powdered.

Step 3: Soaking the Ground Plant Samples in a Solvent. Soaking the ground
samples in a specific solvent will make the compounds go to the solvent or will
dissolve in the solvent. This is already the start of the extraction process. The
kind of solvent you want to use depends on your study particularly on the
specific compound you want to extract from the plants. Most commonly used
are ethanol, acetone, petroleum ether, and hexane.

Step 4: Collecting the filtrate. The dissolved compounds from the plant
samples can be found in the filtrate, the liquid mixture. Filter the mixture using
a filter paper (usually Whatman #1 filter paper or as required by the
procedure).

Step 5: Evaporating the solvent. This is the last step in preparing the crude
compounds of crude extract from the plant sample. You can evaporate the
solvent using a Rotatory Evaporator (Rotavap) or you can simplify by
evaporating the solvent by heating using an alcohol lamp. The crude extract
should be in semi-solid form.

Step 6: Recover the extract from the rotary evaporator container. The
extract should be semi-solid or sticky in appearance to ensure that the solvent
ethanol or other solvent chemicals are thoroughly removed.

3) Titration
This is the process of determining the concentration of a solution. Medwick &
Kirschner (2010) defined titration as “a common laboratory method of
quantitative
chemical analysis that used to determine the unknown concentration of an
identified analyte.” The common types of titration are acid-base titration and
redox titration. An acid-base titration depends on the neutralization between
an acid and a base when mixed in a solution. When there is a change in color in
the acid-base indicator signals the endpoint of the titration.

C. Physical

1) Aeration
 This is the interchange of various gases (CO 2 & O2) between the atmosphere
and soil and the various reactions that either consume or produce gases in the
soil.

For example, the oxygen gas (O 2) moves from the atmosphere to soil and is
consumed by plant roots and microorganisms. While carbon dioxide (CO 2)
moves from soil, where it is produced by the plant and microbial respiration, to
the atmosphere.

2) Centrifugation
This is the process of separating substances applying centrifugal force
through the use of a centrifuge. A centrifuge is a device used to separate
particles or macromolecules (i.e., cells, nucleic acids, proteins, and sub-cellular
components).

3) Chromatography
The word chromatography means “color writing”, which chemists used to test
liquid mixtures. This is the process of separating organic and inorganic
compounds used by scientists to analyze and study these compounds.

There are four main types of chromatography: Gas Chromatography, Liquid

Chromatography, Paper Chromatography, and Thin-layer Chromatography.

The table below presents a detailed comparison of these types.

Types of Chromatography Description Application(s)

- Used to analyze volatile o Detects bombs in
gases. airports
o Identifies and
quantifies drugs, i.e.,
a) Gas Chromatography
in alcohols o
(Forensics)
Compares fibers
found on a victim
- Used to analyze the o Detects pollution by
metal ions and organic testing water
compounds in samples
solutions.
b) Liquid Chromatography - uses liquids that may
incorporate
hydrophilic, insoluble
molecules
- common type of o Separates amino
chromatography acids and anions
- paper is the stationary o RNA fingerprinting o
c) Paper Chromatography phase Separates and tests
- uses capillary action to histamines and
pull the solutes up antibiotics
through the paper and
separate the solutes
 o Detects pesticide or
- simple and quick insecticide residues
d) Thin-layer method to check the in food
Chromatography purity of the organic o Analyzes the dye
compounds composition of fibers
(forensics)

4) Distillation

This is the process by which organic compounds (contain carbon) are purified.
7 This is advantageous in the sense that two different compounds have
different boiling points. Take, for example, two different liquids are present in a
homogeneous mixture (completely miscible, e.g., H 2O & alcohol). If these
compounds have different boiling points, one of the compounds will evaporate
first (volatile) before the other will.

5) Drying

This is defined as the process of vaporizing and removing water or other

liquids in a certain material to form a dry solid. Physiochemical transformations
result from the simultaneous heat and mass transfer (which are critical factors
in the drying process).

6) Grinding

This is the process of turning a material into small pieces from fine to coarse
using a certain device, i.e., Wiley Mill or a grinding wheel.

7) pH measurement
A pH is a measurement of how weak and strong a particular acidic or basic
substance. A substance with a pH of 7 is neutral, i.e., H 2O. A pH lower than 7
means acidic, i.e., acetic acid found in vinegar. A pH higher than 7 is a base,
i.e., sodium hypochlorite found in bleaching products.

In many studies, pH is an important parameter or variable. For example, pH is

an important parameter or factor that is measured to determine the condition
of water because of pollution. How do you relate pH measurement with
pollution or water quality? With the use of pH, one can measure how acidic or
basic water is to determine its water quality. Pure water has a pH of 7, a little
lower or higher than that can be considered normal. But when the pH goes
significantly lower, it means the substance is already acidic and basic when it
goes far beyond pH 7. If drinking water for example becomes acidic or basic
only indicates that other components are visible. A pH of 7.4. of water in rivers
means good quality. Any higher or lower than pH 7 means harm to the
organisms living in the water.

8) Weighing
This procedure shows how o use a balance to make a precise and accurate
measurement, i.e., mass. To do this, the use of measuring devices is very
essential.

Example (Measuring devices)

 o Solids – triple balance, top-pan electric balance, analytical electric
balance, etc.
o Liquids or solids that do not holder together (i.e., powder, ice, etc.) –
receptacle, graduated cylinder, etc.

III. Activity Proper

Activity 2. Experiment @ the Lab

Objective: Identify the standard methods and techniques in performing
experiments.

Directions. Using the given statement/title in each number, try to identify the
laboratory techniques being described. Write your answer on a separate sheet of
paper.

1. A particular food color mixture was separated into two colors – red and blue.

2. The human blood needs to keep its pH approximately 7.4 to maintain stability
to avoid any complications from it.

3. Bioprospecting for Novel Halophilic and Halotolerant Sources of Hydrolytic

Enzymes in Brackish, Saline and Hypersaline Lakes of Romania

4. The tannins, theobromine, and caffeine are obtained from the tea leaves in
boiling water.

5. Alcoholic beverages like whiskey are produced from fermented grains like
corn or rye at distilleries.

6. In baking, a cake of different varieties in yellow, white, and pound cakes to

produce a cake of correct volume, grain, and texture, the amount of air into
the batter needs to be considered.

7. Natural and synthetic food coloring are used as additives to make food more
acceptable and popular. Non-permitted coloring agents are not added to the

10
foods, instead, edible colors are. Thus, these coloring agents are identified and
quantified.

8. Pharmaceutical industries used UV and fluorescence spectrometers invariably

to provide detectors for the instrumental-based systems.

9. A baker wanted to know the amount of salt or sugar that might affect the
color of the cake that she will bake.

10. Removing water in wet lettuce with the use of a salad spinner
 Questions
Directions: Answer the following on a separate sheet of paper.
1. Based on the activities that you have accomplished, why do you need to know
the different laboratory methods and techniques in performing experiments?

2. Among the laboratory methods and/or techniques you have learned, which
would you likely utilize in conducting experiments that would answer your
research questions?

IV. Reflection
Complete the statements below.

I understand_____________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
I don’t understand _____________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
I need more information about
___________________________________________________
_________________________________________________________________________________
_________________________________________________________________________________

V. Links and Other References

Experimental Methods. 2021. ScienceDirect ®. Retrieved on February 9, 2021,
from https://www.open.edu/openlearncreate/mod/oucontent/view.php?
id=233&pri ntable=1

Lahora, Oliver A. (2020). Scientific Research for Junior High School. pp. 45-46, 55,
137

Retrieved on February 9, 2021, from https://eda.nc3rs.org.uk/experimental-

designunit

Retrieved on February 9, 2021, from https://www.pubmed.ncbi.nlm.nih.gov

Retrieved on February 9, 2021, from https://biotopics.co.uk/microbes.tech1.html

Retrieved on February 9, 2021, from https://www.microbiologyonline.org>file

Retrieved on February 9, 2021, from https://www.chemengonline.com/solidsdrying-
basics-and-applications/?printmode=1

Retrieved on February 9, 2021, from https://www.google.com/amp/s/dictionary

.comcambridge.org/us/

Retrieved on February 9, 2021, from

https://www.sciencedirect.com/topics/agricultural-and-biologicalsciences/
titration

Retrieved on February 9, 2021, from

https://chem.libretexts.org/Bookshelves/Organic_Chemistry/Book%3A_Organ
ic_Chemistry_Lab_Techniques_(Nichols)/

Questions