SELF-LEARNING PACKAGE IN
BIOTECHNOLOGY
RHODA M. ERDAO. PhDBy:
Stephanie N. Sanonte
Oton NHS - SDO Iloilo
SELF-LEARNING PACKAGE IN
BIOTECHNOLOGY
Laboratory Apparatus
Learning Competency:
Discuss the proper ways in
handling the Autoclave, Balance,
Pipette and Buret
Ready to Launch!
When you think of Biotechnology, you probably imagine them in a modern
laboratory with test tubes, other delicate instruments, apparatus, and bottles of
strange substances and materials.
You’re right! In the laboratory, you can find different laboratory apparatus.
The Science laboratory is a place of adventure and discovery. Some of the most
exciting events in scientific history have happened in the laboratory. The discov-
ery of the atoms, the analysis of chemicals of substances, the discovery of the
cell and other first discoveries were done in a laboratory. But all these things
could never have happened if there were no equipment and devices. Different
laboratory apparatus served their purposes. Here are some of the laboratory
equipment and their uses.
Aim at the Target!
After going through this module, you are expected to:
1. discuss the proper ways in handling the autoclave, balance, pipette
and buret
Try This!
Can you still recall some apparatus found inside the laboratory? Identify
and name each of them by choosing your answer inside the box.
Autoclave Double-pan balance Triple beam balance Buret Pipette
1.
2. 3. 4.
Keep This in Mind!
Name ____________________________________________ Score ______________
Year & Section ___________________________________ Date _______________
Activity 1.1– The “Autoclave’s way”
You can visit this website: https://microbenotes.com/autoclave/
Direction: Identify the different parts of an autoclave by choosing your
answer inside the box.
Handles Outer stand Vacuum Release valve Autoclave body Autoclave lid
Steam release valve Pressure regulating device Safety valve Pressure gauge
1. 2. 3.
4.
5.
6.
7.
8.
9.
Keep This in Mind!
Activity 1.2- The “Autoclave’s way” (Procedure)
You CAN visit this website: https://microbenotes.com/autoclave/
Direction: Arrange the following procedures in running an autoclave by
writing a letter before each number.
_____1. The drainage pipe is then closed, and the steam inside is allowed
to reach the desired levels (15 lbs in most cases).
_____2. Now, the electric heater is switched off, and the autoclave is al-
lowed to cool until the pressure gauge indicates the pressure inside has low-
ered down to that of the atmospheric pressure.
_____3. Finally, the lid is opened, and the sterilized materials are taken
out of the chamber.
_____4. Now, the materials to be sterilized are placed inside the chamber.
_____5. Before beginning to use the autoclave, it should be checked for
any items left from the previous cycle.
_____6. The safety valves are adjusted to maintain the required pressure
in the chamber.
_____7. After the whistle, the autoclave is run for a holding period, which
is 15 minutes in most cases.
_____8. A sufficient amount of water is then put inside the chamber.
_____9. The discharge pipe is then opened to allow the entry of air from
the outside into the autoclave.
_____10. Once the pressure is reached, the whistle blows to remove ex-
cess pressure from the chamber.
_____11. The lid is then closed, and the screws are tightened to ensure an
airtight condition, and the electric heater is switched on.
_____12. Once the water inside the chamber boils, the air-water mixture
is allowed to escape through the discharge tube to let all the air inside to be
displaced. The complete displacement can be ensured once the water bubbles
cease to come out from the pipe.
Keep This in Mind!
Activity 2: “Let’s Bu-READ”
A buret is a piece of volumetric glassware used to deliver variable liquid
amounts typically for titrations. It is a long glass cylinder open at one end and
fitted at the opposite end with a stopcock valve (Figure 1). The stopcock valve
controls the flow of liquid from the cylinder through the buret tip. The buret
used in the General Chemistry lab has a 50-mL volume and a Teflon stopcock
requiring no lubrication. The buret is etched with calibration markings at 0.10-
mL intervals allowing the estimation of variable liquid levels to the hundredths
of a milliliter (0.00 mL). Record all numbers from markings and one number
estimated between markings (significant figures) to indicate the sensitivity of
the volume measurement.
Procedure:
Read and write the liquid level contained in the buret.
Answer: _______________
Figure 1. Buret Use.
Keep This in Mind!
Direction: Answer each question.
1. Why are the sides of the buret tapped after filling it?
2. Why are a few milliliters of liquid drained through the tip after filling the buret?
3. After delivering a certain volume into a flask, a drop of liquid is hanging from
the tip. Is it part of the delivered volume? Should it be added to the flask?
Activity 3: “The Heavy Weight is Over”
Direction: Follow the given procedures below.
1. You will pick an object from your possessions to weigh. List the objects chosen
and your estimation of their masses in the table.
Object Estimated Mass Actual Mass
Table 1: Masses of the objects
2. You have to determine the weight of the objects using the balance and fill in the
actual mass column of the previous table.
3. Explain your procedure for finding the mass of your object. Include all calcula-
tions. You may use drawings to assist your explanation.
Abstraction and Generalization
An autoclave is a machine that provides a physical method of steriliza-
tion by killing bacteria, viruses, and even spores present in the material put
inside of the vessel using steam under pressure.
• Autoclave sterilizes the materials by heating them up to a particular tem-
perature for a specific period of time.
• The autoclave is also called a steam sterilizer that is commonly used in
healthcare facilities and industries for various purposes.
• The autoclave is considered a more effective method of sterilization as it is
based on moist heat sterilization.
Autoclave Parts/ Components
The simplest form of the autoclave is the pressure cooker types or labor-
atory bench autoclaves. The following is the detailed description of different
components/ parts of an autoclave:
a. Pressure Chamber
• The pressure chamber is the
main component of a steam au-
toclave consisting of an inner
chamber and an outer jacket.
• The inner chamber is made up of
stainless steel or gunmetal,
which is present inside the out
chamber made up of an iron
case.
• The autoclaves used in
healthcare laboratories have an
outer jacket that is filled with
steam to reduce the time taken
to reach the sterilization temperature. Figure 1: Autoclave
• The inner chamber is the case where the materials to be sterilized are put.
• The size of the pressure chamber ranges from 100 L to 3000 L.
b. Lid/ Door
• The next important component of an autoclave is the lid or door of the au-
toclave.
• The purpose of the lid is to seal off the outside the atmosphere and create
a sterilized condition on hot inside of the autoclave.
• The lid is made airtight via the screw clamps and asbestos washer.
• The lid consists of various other components like:
Pressure gauge
• A pressure gauge is present on the lid of the autoclave to indicate
the pressure created in the autoclave during sterilization.
• The pressure gauge is essential as it assures the safety of the auto
clave and the working condition of the operation.
Pressure releasing unit/ Whistle
• A whistle is present on the lid of the autoclave is the same as that of
the pressure cooker.
• The whistle controls the pressure inside the chamber by releasing a
certain amount of vapor by lifting itself.
Safety valve
• A safety valve is present on the lid of autoclave, which is crucial in
cases where the autoclave fails to perform its action or the pressure
inside increases uncontrollably.
• The valve has a thin layer of rubber that bursts itself to release the
pressure and to avoid the danger of explosion.
c. Steam generator/ Electrical heater
• An electrical steam generator or boiler is present underneath the
chamber that uses an electric heating system to heat the water and
generate steam in the inner and the outer chamber.
• The level of water present in the inner chamber is vital as if the water
is not sufficient; there are chances of the burning of the heating sys-
tem.
• Similarly, if the water is more than necessary, it might interfere with
the trays and other components present inside the chamber.
d. Wastewater cooler
• Many autoclaves are provided with a system to cool the effluent before
it enters the draining pipes.
• This system prevents any damage to the drainage pipe due to the boil-
ing water being sent out of the autoclave.
Procedure for running an autoclave
YOU may watch this video: https://www.youtube.com/watch?v=0dlDf8HiiQw&feature=emb_logo
In general, an autoclave is run at a temperature of 121° C for at least 30
minutes by using saturated steam under at least 15 psi of pressure. The follow-
ing are the steps to be followed while running an autoclave:
1. Before beginning to use the autoclave, it should be checked for any items left
from the previous cycle.
2. A sufficient amount of water is then put inside the chamber.
3. Now, the materials to be sterilized are placed inside the chamber.
4. The lid is then closed, and the screws are tightened to ensure an airtight con-
dition, and the electric heater is switched on.
5. The safety valves are adjusted to maintain the required pressure in the
chamber.
6. Once the water inside the chamber boils, the air-water mixture is allowed to
escape through the discharge tube to let all the air inside to be displaced. The
complete displacement can be ensured once the water bubbles cease to come
out from the pipe.
7. The drainage pipe is then closed, and the steam inside is allowed to reach the
desired levels (15 lbs in most cases).
8. Once the pressure is reached, the whistle blows to remove excess pressure
from the chamber.
9. After the whistle, the autoclave is run for a holding period, which is 15
minutes in most cases.
10. Now, the electric heater is switched off, and the autoclave is allowed to cool
until the pressure gauge indicates the pressure inside has lowered down to that
of the atmospheric pressure.
11. The discharge pipe is then opened to allow the entry of air from the outside
into the autoclave.
12. Finally, the lid is opened, and the sterilized materials are taken out of the
chamber.
BURETS
You may visit this site: (RDGburets-file:///C:Users/Teacher/Desktop/RDGburets.pdf)
A buret is a piece of volumetric glass-
ware used to deliver variable liquid amounts
typically for titrations. It is a long glass cylin-
der open at one end and fitted at the oppo-
site end with a stopcock valve. The stopcock
valve controls the flow of liquid from the cyl-
inder through the buret tip.
The numbering of the markings begins
at the top (open end) of the buret. Therefore,
the volume delivered can be determined from
the difference between initial and final liquid
levels. Refer to Figure 2 and note the correct
way to read the liquid level in a buret (from
the top down).
Before use, a buret should always be
cleaned and prerinsed with the solution to be de- Figure 2: Buret use.
livered. Do NOT attempt to fill a buret precisely to
the 0.00-mL mark at the top. This is a waste of time, as all volumes delivered by
a buret are determined from the difference between initial and final readings.
After filling the buret with solution, drain a few milliliters of liquid through the
stopcock into a beaker to remove any air bubbles trapped in the valve. Close the
stopcock, tap the buret sides gently, and allow the liquid to stand for a few
minutes to clear gas bubbles dissolved in the liquid. Remove any drop hanging
from the buret tip by touching the tip to the side of the beaker or wiping the tip
with a tissue. Read the initial volume to the hundredths place (0.00 mL).
To transfer the liquid, open the stopcock valve until the desired amount of
liquid is slowly drained into a container. Close the valve and touch off the hang-
ing drop of liquid on the buret tip adding it to the liquid in the container. This
step is important because the hanging drop is part of the volume delivered by
the buret. Allow a moment for liquid on the cylinder walls to settle before re-
cording the final volume to ±0.01 mL (again from the top down). The difference
between the initial and final volumes is the amount of liquid transferred to the
container.
Buret Care
• Wash the buret with a mild dilute soap solution. Rinse first with tap water,
and then deionized (DI) water. If beads of water form on the walls, rewash.
• Never dry a buret in an oven! The heat will distort the glass and change the
calibrated volumes. Never dry a buret using air jets! The air system contains
oil droplets and fine dust, also the compressed air can break the glass.
• A buret should always be rinsed with a small amount of the solution to be
used. This step prevents contamination or dilution of the solution from wa-
ter or other chemicals on the glassware’s inside walls and removes the need
to dry the buret.
• After an experiment is completed, drain all chemicals from glassware, pour-
ing them into marked collection containers in the hoods. Glassware may be-
come irreversibly etched from prolonged contact with some solutions
(especially NaOH or KOH solutions). Wash the glassware, rinse thoroughly,
and return to common storage area at the front of the laboratory.
• Return all broken or cracked glassware to the stockroom for replacement.
Triple Beam Balance
You may watch this video: https://www.youtube.com/watch?v=l5q_bwiAdCc
A balance is generally used for weighing out small amounts of chemicals
to use in solutions and determining the mass of different objects. Learning how
to use a balance is important for collecting data or information and for ensuring
accurate mass measurements.
Figure 3: Triple Beam Balance
Special Safety, Care, and Handling Aspects
• Make sure the riders are at zero before carrying the balance.
• Carry the balance with both hands and keep it horizontal.
• Do not place anything hot directly on the balance pan.
• Never pour chemicals directly on the balance pan.
• Place balance on a level, sturdy surface and away from heavy traffic.
Setting Up the Balance
• Make sure the riders are at zero.
• Check to make sure the pointer swings freely along the scale.
• Use the adjustment screw to obtain an equal swing of the beams, if neces-
sary.
Pipettes
A pipette is a small tube that can transfer liquids from one container
to another. Pipettes, or pipets, are common pieces of laboratory equip-
ment.
Pipettes may be constructed out of glass or plastic and are used to
transfer a measurable amount of liquid.
They are designed either to contain (TC) or to deliver (TD) a specific
volume and will be stamped as such by the manufacturer. TC pipettes
will contain a specified volume but will not dispense the exact volume,
and TD will dispense the exact volume.
Types of Pipettes
1. Volumetric pipettes transfer a single, predetermined
volume of liquid. They are often called a bulb pipette
for their shape, which is a long tube-like shaft at the
bottom and top and a bulb in the center where the
bulk of solution is held.
2. Measuring Pipettes and Micropipettes:
Measuring pipettes have graduated volumes, like
a graduated cylinder, and can dispense different vol-
umes. Micropipettes are accurate and precise and can
transfer measured volumes of microliters of volume.
Reflect
Think, Write, Draw and Share!
Laboratory apparatus are very useful in almost all of our science activities.
Given that you are already knowledgeable of the different apparatus and their
functions. Be imaginative enough to draw your dream Science lab. Place this
in a short bond paper. I’ve got mine below.
_____________________________________________________________________
_____________________________________________________________________
Reinforcement & Enrichment
Web Search!
1. In a short-sized bond paper with portrait orientation, search,
copy and paste new articles, researches or innovations done in laboratories
about CORONA VIRUS.
2. This output will be submitted at the end of the week.
3. Happy working !
Assess Your Learning
I. Direction: Read and understand each item carefully. Write the letter
of the correct answer.
____1. A machine that provides a physical method of sterilization by killing bac-
teria, viruses, and even spores present in the material put inside of the vessel
using steam under pressure.
a. Buret b. Pipette c. Autoclave d. Triple beam balance
____2. A small tube that can transfer liquids from one container to another.
a. Double-pan balance b. Autoclave c. Buret d. Pipette
____3. It is present on the lid of the autoclave to indicate the pressure created in
the autoclave during sterilization.
a. Pressure gauge b. Safety valve c. Whistle d. Steam generator
____4. It is generally used for weighing out small amounts of chemicals to use in
solutions and determining the mass of different objects.
a. Pipette b. Balance c. Buret d. Autoclave
____5. It is a long glass cylinder open at one end and fitted at the opposite end
with a stopcock valve.
a. Autoclave b. Buret c. Balance d. Pipette
II. Direction: True or False: Write T if the statement is correct and F if other-
wise.
_____1. The pressure chamber is the main component of a steam autoclave con-
sisting of an inner chamber and an outer jacket.
_____2. A pressure gauge is present on the lid of autoclave, which is crucial in
cases where the autoclave fails to perform its action or the pressure inside in-
creases uncontrollably.
_____3. An electrical steam generator or boiler is present underneath the cham-
ber that uses an electric heating system to heat the water and generate steam
in the inner and the outer chamber.
____4. The correct way to read the liquid level in a buret is from the top down.
____5. In setting up a triple beam balance, make sure the riders are at 1 or
pointing in 1.
References & Photo Credits
https://courses.lumenlearning.com/ap1x94x1/chapter/the-parts-of-a-compound-
microscope-and-how-to-handle-them-correctly/
https://www.youtube.com/watch?v=pujis3QsA_I
https://lrmds.deped.gov.ph/detail/7414
https://lrmds.deped.gov.ph/detail/7413
file:///C:/Users/User/Downloads/Chem%20M2%20Laboratory%20Apparatus,%
20Safety%20Rules%20&%20Symbols.pdf
https://lrmds.deped.gov.ph/detail/7164
https://camblab.info/how-to-pipette-the-essential-guide-to-good-technique/
https://chem.libretexts.org/Bookshelves/Ancillary_Materials/
Demos_Techniques_and_Experiments/General_Lab_Techniques/Proper_Use_of_a_Buret
https://microbenotes.com/autoclave/
https://www.youtube.com/watch?v=Asg55fFt4zk
http://faculty.sites.uci.edu/chem1l/files/2013/11/RDGburets.pdf
https://kscorn.com/wp-content/uploads/2019/11/pipette-technique-2018-Final-
web.pptx
https://www.google.com/search?
q=triple+beam+balance+weighing&rlz=1C1OKWM_enPH921PH921&sxsrf=ALeKk01uNqfar
JhJxQK3DHn1g9zkFmxDKg:1602723191307&source=lnms&tbm=isch&sa=X&ved=2ahUK
EwjIm_GTsbXsAhXZMd4KHRPtBk0Q_AUoAXoECBEQAw#imgrc=RmU-lx0dHcR5KM
https://www.youtube.com/watch?v=l5q_bwiAdCc
https://sciencing.com/purpose-pipette-8743073.html
https://labproinc.com/collections/volumetric-pipettes
https://www.nichiryo.co.jp/en/productline/pipette.html
https://stevenlie.blogspot.com/2009/11/futuristic-science-lab.html
ANSWERS
TRY THIS!
1. Triple beam balance
2. Autoclave
3. Buret
4. Pipette
Activity 1.1– The “Autoclave’s way”
1. Pressure regulating device
2. Pressure gauge
3. Safety valve
4. Autoclave lid
5. Handles
6. Autoclaves body
7. Steam release valve
8. Vacuum release valve
9. Outer stand
Activity 1.2- The “Autoclave’s way” (Procedure)
1. G
2. J
3. L
4. C
5. A
6. E
7. I
8. B
9. K
10. H
11. D
12. F
ANSWERS
Activity 2: “Let’s Bu-READ”
Answer: 9.36 mL
1. To allow the liquid to stand for a few minutes and to clear gas bubbles dissolved in the water.
2. After filling the buret with solution, drain a few milliliters of liquid through the stopcock into a
beaker to remove any air bubbles trapped in the valve.
3. NO. NO.
Activity 3: “The Heavy Weight is Over”
Answers may vary.
Assess Your Learning
I. Multiple Choice
1. C
2. D
3. A
4. B
5. B
II. True or False
1. T
2. F
3. T
4. T
5. F
