MOLECULAR BIOLOGY & DIAGNOSTICS
P-LAB1: BASIC LABORATORY PRACTICES; MOLBIO
LAB DESIGN, EQUIPMENT, & INSTRUMENTS
Transcriber: Langit, John Bern R. | Professor: Prof. Adlene M. Atienza
1.0 BASIC LABORATORY PRACTICES
2.0 MOLBIO LABORATORY DESIGN
General Laboratory Practices:
1) Always wear appropriate personal protective equipment
(PPE).
a. Lab coats should be worn in the lab to protect you and
your clothing from contamination. Lab coats should
not be worn outside the laboratory.
b. Lab footwear should consist of normal closed shoes to
protect all areas of the foot from possible puncture
from sharp objects and/or broken glass and from
contamination from corrosive reagents and/or
infectious materials.
c. Gloves should be worn, and change gloves when
contaminated.
d. Protective eyewear and/or masks may need to be worn
when contact with hazardous aerosols, caustic
chemicals and/or reagent is anticipated.
2) Hands are washed after removing the gloves, before and after
completion of work.
3) Do not eat, drink, smoke, handle contact lenses, apply
cosmetics, or store food for human consumption in the
laboratory.
4) Safely handle sharps and place them on puncture-proof
containers after use. Used needles are not removed from
disposable syringes nor bent or recapped.
5) No mouth pipetting.
6) Take care to minimize the creation of aerosols and/or splashes.
Use closed containers.
7) Manipulations of infectious materials must be conducted in
the biosafety cabinet.
8) Decontaminate all work surfaces before and after your
experiments/work, and immediately after any spill or splash of
potentially infectious material with a 10% bleach.
9) Decontaminate all potentially infectious materials before
disposal. Biohazardous material should be discarded in a
biohazard bag to be autoclaved before disposal.
10) Report any incidents that may result in exposure to infectious
materials to appropriate personnel (e.g. Laboratory supervisor,
safety officer).
Standard Requirements of COVID-19 Testing Laboratories:
Source: DOH Administrative Order No. 2020-0014, Guidelines in Securing a License to
Operate a COVID-19 Testing Laboratory in the Philippines
•
Other Guidelines for Molecular Biology Laboratory:
― Emergency equipment
― Ultraviolet light
― Electricity
― General Housekeeping
― Preparing for the lab
― Following protocols
― Sterility
― Hazards in the lab
― Equipment and reagents
Physical Facilities: COVID-19 testing laboratories shall have
adequate and appropriate areas to safely, effectively and
efficiently provide the services to clients.
1)
2)
3)
4)
5)
Prototype Floor Plan:
COVID-19 Testing Laboratory using RT-PCR
•
•
•
Page 1 of 15
There shall be a dedicated space for each of the
following activities:
a. Specimen reception;
b. Virus inactivation and nucleic acid
extraction (Pre-PCR);
c. Reagent storage and handling;
d. PCR; AND
e. Clerical activities.
Unidirectional workflow following the
abovementioned activities shall be maintained at all
times.
The prototype floor plan and the floor plan checklist
for constructing a COVID-19 testing laboratory shall
be used as references in constructing the testing
laboratory. (ANNEX D1 and ANNEX D2).
Controlled and adequate ventilation with the
prescribed air changes per hour shall be maintained
for each specific area.
Adequate lighting shall be provided in all areas.
Ante-Room: specific area designed to maintain the
pressure differential between the laboratory and its
adjacent space
Positive Pressure: airflow outward
― No contaminated air will go in.
― Example: Reagent Preparation Room
Negative Pressure: airflow inward
― No contaminated air will go out.
― Example: Sample Preparation and
Amplification Rooms; Isolation Rooms
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
Features of Molecular Biology Laboratory Design:
LANGIT 2023
3.0 MOLBIO LABORATORY EQUIPMENT & INSTRUMENT
Laboratory Equipment and Instruments:
Molecular Biology Laboratory Design
1.
2.
3.
4.
5.
6.
7.
8.
9.
Autoclave
Biological Safety Cabinet (BSC)
Polymerase Chain Reaction Machine
Spectrophotometer
Vortex Mixer
Centrifuge
Automatic Pipettors
Refrigerator
pH Meter
(1) Autoclave:
•
•
•
•
It consists of three areas, namely the:
a. Reagent Preparation Room/Area
b. Sample Preparation Room/Area
c. Amplification Room/Area
It operates in a unidirectional workflow.
o The sequence starts from: (a) Reagent Preparation, (b)
Sample Preparation, to (c) Amplification.
o There must be no backflow traffic. It must be
restricted to minimum!
Its areas operate in a particular airflow.
 Reagent Preparation Area → Positive Pressure
 Sample Preparation Area → Negative Pressure
 Amplification Area
→ Negative Pressure
Its areas consist of processes, some are in a particular order
yet some are not.
a. Reagent Preparation Area:
•
Reagent Storage
•
Reagent Preparation
b. Sample Preparation Area:
1. Specimen Preparation
2. Nucleic Acid Isolation
c. Amplification Area:
1. Amplification
2. Detection Analysis

It provides a physical
method of sterilization by
killing bacteria, viruses,
and even spores present in
the material put inside of
the vessel using steam
under pressure (aka heat).

What makes the autoclave efficient?
― For efficiency, maintain 121 degrees Celsius within
20-30 minutes at 15 psi pressure.
Materials to be autoclaved depend on the ff factors:
a. Time
b. Pressure
c. Steam
d. Quantity


Materials such as:
― Plastic (shorter time and pressure)
― Glass (longer time and pressure)
Types of Autoclaves:
1. Pressure Cooker Type
2. Common Laboratory Autoclave
3. Vertical Autoclave
4. Horizontal Autoclave
5. Large Automatic Hospital Autoclave
Molecular Biology Laboratory in FEU-NRMF
Page 2 of 15
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
LANGIT 2023
(2) Biological Safety Cabinet (BSC):
Class 1 BSC:
 It is the first designed and simple Biological Safety
Cabinet (BSC) which provides personnel and
environmental protection but not product production (as
unsterilized room air is drawn over the work surface).
 These are suitable for work with Risk Group 1 (RG1),
Risk Group 2 (RG2), and Risk Group 3 (RG3) biological
material.

It is an engineering
control intended to
protect laboratory
workers, laboratory
environment and work
materials from exposure
to infectious or
biohazardous aerosols
and splashes.
―
Such aerosols and splashes may be generated while
manipulating materials containing infectious agents, such as
primary cultures, stocks and diagnostic specimens.


Factors that produce biohazard aerosols:
― Rapid hand movements
―

What makes the BSC efficient?
― HEPA Filters
HEPA Filters:
 High Efficiency Particulate Air (HEPA) Filters
 They are constructed of pleated borosilicate glass and
arranged into random fibers.
 It traps 99.97% of particles of 0.3 µm in diameter (anything
larger to 0.3 can be efficiently filtered by this filter) since
gases and vapors are relatively smaller they cannot be
removed by this filtration.
Working Mechanism:
― Room air is drawn in through the front opening at a
minimum velocity of 0.38 m/s.
― The front opening also allows the operator’s arms
to reach the work surface inside the cabinet while
he or she observes the work surface through a glass
window.
― The window can also be fully raised to provide
access to the work surface for cleaning or other
purposes.
― The directional flow of air whisks aerosol particles
that may be generated on the work surface away
from the laboratory worker and are then discharged
from the BSC through a HEPA filter.
HEPA filter traps 99.9% of particles of ≥ 0.3 µm in diameter.
This enables the HEPA filter to effectively trap all known
infectious agents and ensure that only microbe-free exhaust air is
discharged from the cabinet and/or recirculated in the work
surface.
Class II BSC:
 It is a ventilated cabinet, which provides personnel, product
and environmental protection.
 It is commonly found in clinical and research laboratories
working with infectious agents in Risk Groups 2, 3 and 4
(if positive-pressure suits are used) or with tissue culture.
•
Page 3 of 15
There are four types: A1, A2, B1, and B3. The main
differences between the types are the ratio of air exhausted
from the BSC to the air that is recirculated within the BSC,
and the type of exhaust system present.
o About 90% of all biosafety cabinets installed are
Type A2 cabinets. There is a limited need for
Class II Type B biological safety cabinets.
o In addition, Class II Type B biological safety
cabinets require very specific installation and
operating conditions to function correctly.
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS


Working Mechanism:
― The working mechanism of Class II BSCs differs
according to their types.
― Protection Features:
o It has an open front with inward airflow
for personnel protection,
o downward HEPA filtered laminar airflow
over the work surface for product
protection and
o HEPA filtered exhausted air for
environmental protection.
― The room air and recirculated air are HEPA filtered
before flowing downwards over the work area.
― Class II BSCs can be exhausted into the containment
zone or directly to the outside atmosphere through a
thimble or hard-ducted connection depending on the
type.
― The amount of air that recirculates or exhausts
depends on the type.
Class III BSC:
 It provides the highest level of personnel protection and is
used for Risk Group 4 agents. It is suitable for work in
Biosafety Level 3 and 4 laboratories.
LANGIT 2023
Working Mechanism:
― This type of cabinet is totally enclosed and is tested
under pressure to ensure that no particles can leak
from it into the room.
― Supply air is HEPA-filtered and exhaust air is
discharged into the atmosphere through two HEPA
filters.
― The operator access the work surface by means of
heavy-duty rubber gloves which form part of the
cabinet. Several glove boxes can be joined together to
extend the work surface.
― Airflow is maintained by a dedicated exhaust system
exterior to the cabinet, which keeps the cabinet
interior under negative pressure.
― The Class III BSCs should have an attached passthrough box that is sterilizable and equipped with
HEPA-filtered exhaust.
― The Class III cabinet may be connected to a doubledoor autoclave used to decontaminate all materials
entering or exiting the cabinet.
Table: Biological Safety Cabinet (BSC) Classes
Protection
Biosafety
Risk Group
Level
Agent
Laboratory
Class I
- Personnel
- RG1
- Environment
- RG2
- RG3
Class II - Personnel
- Clinical and
- RG2
- Environment
Research
- RG3
- Product
Laboratories
- RG4 (if
positive
pressure suits
are used)
- Tissue
Culture
Class
- Personnel
- Level 3
- RG4
III
- Environment
- Level 4
- Product
Class
Type
Class I
Class
IIA1
Class
IIA2
Class
IIB1
Class
IIB2
Class
III
Page 4 of 15
Table: Features of Class I, II, and III BSCs
Face
Airflow
Airflow
Exhaust
Velocity (recirculated) (exhausted) System
(m/s)
0.36
0
100
Hard duct
0.3870
30
Exhaust to
0.51
room or
thimble
connection
0.51
70
30
Exhaust to
room or
thimble
connection
0.51
30
70
Hard duct
0.51
0
100
Hard duct
NA
0
100
Hard duct
BSMT 2, SEMESTER 2
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
(3) Polymerase Chain Reaction Machine:

1ST PRELIMINARY TERM
(5) Vortex Mixer:

laboratory technique
used to make multiple
copies of a segment of
DNA
Conventional PCR
simple device used
commonly in laboratories
to mix small vials of
liquid
Real Time PCR
(6) Centrifuge:

Centrifugation is a
procedure that separates
components of liquids
that have different
weights. The device used
for this purpose is called a
centrifuge, in which a
centrifugal force is
applied that moves liquid
components away from
the center.
(7) Automatic Pipettors:

(4) Spectrophotometer:

is an instrument used to
measure how much a
chemical substance
absorbs light by
measuring the intensity
of light as a beam of light
passes through sample
solution.
Pipettes are equipment
used to deliver an
accurately measured
volume of solution. In a
molecular biology
laboratory,
micropipettes are most
frequently used.
Micropipettes measure
volumes that range from
1000 μl (1 ml) to less
than 1 μl.
(8) Refrigerator:
Page 5 of 15
LANGIT 2023
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
(9) pH Meter:

very sensitive to
changes in pH and
hence, buffers are
used to stabilize
the pH
REFERENCES
•
•
PPT Handout (Prof. Atienza)
Administrative Order No. 2020-0014: Guidelines in Securing
a License to Operate a COVID-19 Testing Laboratory in the
Philippines (https://doh.gov.ph/sites/default/files/healthupdate/ao2020-0014.pdf)
o Article V Implementing Mechanisms, Letter B
Specific Guidelines, Number 2 Standards, Letter B
Physical Activities
o Annex D1-D2
PPT Handout References:
•
•
•
•
•
•
•
•
•
Laboratory biosafety manual, Third edition, World Health
Organization (WHO), 2004.A Guide to Biosafety &
Biological Safety Cabinets. ESCD.
Richmond JY, McKinney RW. Primary containment for
biohazards: selection, installation and use of biological safety
cabinets, 2nd ed. Washington, DC, U.S. Department of Health
and Human Services/Centers for Disease Control and
Prevention/National Institutes ofHealth, 2000.
Health Canada. Laboratory biosafety manual, 2nd ed. Ottawa,
Minister of Supply and Services Canada,1996.
Massachusetts Institute of Technology, Environment, Health
& Safety Office, Biosafety Cabinets.
Pelt-Veíkuil EV, van Belkum A, Hays JP. Píinciples and
ľechnical Aspects of PCR Amplification. Spíingeí Netheílands
Publishing, 2008. ISBN: 978-1-4020-6240-7 (Píint) 978-14020-6241-4 (Online).
Collecting, píeseíving and shipping specimens foí the
diagnosis of avian influenza A (H5N1) viíus infection. Guide
foí field opeíations. Woíld Health Oíganization (Inteínet).
Cited 2014 Sep.
http://www.who.int/csí/publications/suíveillance/Annex7.pdf.
https://www.microlit.us/micropipette-product-guide/
https://www.aphl.org/programs/newborn_screening/Document
s/2016%20Molecular%20Workshop/4.2%20%20lab%20design%20QA%20QC%20considerations.pdf
ADDITIONAL DOCUMENTS

Annex D1-D2: Administrative Order No. 2020-0014:
Guidelines in Securing a License to Operate a COVID-19
Testing Laboratory in the Philippines
(https://doh.gov.ph/sites/default/files/health-update/ao20200014.pdf)
Page 6 of 15
LANGIT 2023
BSMT 2, SEMESTER 2
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
1ST PRELIMINARY TERM
Page 7 of 15
LANGIT 2023
BSMT 2, SEMESTER 2
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
1ST PRELIMINARY TERM
Page 8 of 15
LANGIT 2023
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
―
RECALL QUESTIONS
1.
Match:
a.
b.
c.
d.
e.
f.
g.
h.
i.
j.
k.
l.
m.
n.
o.
p.
q.
―
―
―
―
―
―
―
―
―
―
―
―
―
―
―
―
―
―
Ante-Room
Positive Pressure
Negative Pressure
Autoclave
Biological Safety Cabinet (BSC)
Polymerase Chain Reaction Machine
Spectrophotometer
Vortex Mixer
Centrifugation
Automatic Pipettors
pH Meter
micropipettes
Centrifuge
HEPA Filters
Class 1 BSC
Class II BSC
Class III BSC
―
―
―
―
―
LANGIT 2023
It is commonly found in clinical and research
laboratories working with infectious agents in Risk
Groups 2, 3 and 4 (if positive-pressure suits are
used) or with tissue culture.
is a procedure that separates components of liquids
that have different weights.
It provides the highest level of personnel protection
and is used for Risk Group 4 agents. It is suitable for
work in Biosafety Level 3 and 4 laboratories.
It is a ventilated cabinet, which provides personnel,
product and environmental protection.
very sensitive to changes in pH and hence, buffers
are used to stabilize the pH
simple device used commonly in laboratories to mix
small vials of liquid
Answer: OFANJ BLONC GNMEC BLDPI QPKH
1.0 BASIC LABORATORY PRACTICES
These are suitable for work with Risk Group 1
(RG1), Risk Group 2 (RG2), and Risk Group 3
(RG3) biological material.
laboratory technique used to make multiple copies
of a segment of DNA
specific area designed to maintain the pressure
differential between the laboratory and its adjacent
space
They are constructed of pleated borosilicate glass
and arranged into random fibers.
are equipment used to deliver an accurately
measured volume of solution.
No contaminated air will go in.
In a molecular biology laboratory, these are most
frequently used.
It is the first designed and simple Biological Safety
Cabinet (BSC) which provides personnel and
environmental protection but not product production
(as unsterilized room air is drawn over the work
surface).
It traps 99.97% of particles of 0.3 µm in diameter
(anything larger to 0.3 can be efficiently filtered by
this filter) since gases and vapors are relatively
smaller they cannot be removed by this filtration.
No contaminated air will go out.
is an instrument used to measure how much a
chemical substance absorbs light by measuring the
intensity of light as a beam of light passes through
sample solution
High Efficiency Particulate Air (HEPA) Filters
device in which a centrifugal force is applied that
moves liquid components away from the center
It is an engineering control intended to protect
laboratory workers, laboratory environment and
work materials from exposure to infectious or
biohazardous aerosols and splashes.
airflow inward
airflow outward
These measure volumes that range from 1000 μl (1
ml) to less than 1 μl.
It provides a physical method of sterilization by
killing bacteria, viruses, and even spores present in
the material put inside of the vessel using steam
under pressure (aka heat).
2.
General Laboratory Practices:
a. Always wear appropriate ___.
i.
ii.
iii.
iv.
b.
c.
d.
e.
f.
g.
h.
i.
j.
3.
Page 9 of 15
Lab ___ should be worn in the lab to protect
___ and your ___ from ___. These should not
be worn ___ the laboratory.
Lab ___ should consist of normal ___ shoes to
protect all areas of the ___ from possible ___
from sharp objects and/or broken glass and
from ___ from corrosive reagents and/or
infectious materials.
___ should be worn, and change gloves when
___.
Protective ___ and/or ___ may need to be worn
when contact with hazardous aerosols, caustic
chemicals and/or reagent is anticipated.
Hands are washed ___ removing the gloves, ___ and
___ completion of work.
Do not eat, drink, smoke, handle contact lenses,
apply cosmetics, or store food for human
consumption in the laboratory.
Safely handle ___ and place them on ___-proof
containers after use. Used needles are not ___ from
disposable ___ nor ___ or ___.
No ___ pipetting.
Take care to minimize the creation of ___ and/or
___. Use closed containers.
Manipulations of infectious materials must be
conducted in the ___.
___ all work surfaces ___ and ___ your
experiments/work, and immediately after any spill
or splash of potentially infectious material with a
___% ___. (or ___% ___).
___ all potentially infectious materials before
disposal. Biohazardous material should be discarded
in a ___ to be ___ before disposal.
Report any ___ that may result in exposure to
infectious materials to appropriate ___ (e.g.
Laboratory supervisor, safety officer).
Other Guidelines for Molecular Biology Laboratory:
a.
b.
c.
d.
e.
f.
g.
h.
i.
___ equipment
___ light
Electricity
General ___
___ for the lab
Following ___
Sterility
___ in the lab
Equipment and ___
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
2.0 MOL BIO LABORATORY DESIGN
8.
Positive Pressure:
a. airflow ___
b. No contaminated air will go ___.
c. Example/s:
9.
Negative Pressure:
a. airflow ___
b. No contaminated air will go ___.
c. Example/s:
Standard Requirements of COVID-19 Testing Laboratories:
4.
Physical Facilities: COVID-19 testing laboratories shall have
___ and ___ areas to ___, ___ and ___ provide the services to
clients.
1)
2)
3)
4)
5)
5.
There shall be a dedicated ___ for each of the
following activities:
a. Specimen reception;
b. Virus inactivation and nucleic acid
extraction (Pre-PCR);
c. Reagent storage and handling;
d. PCR; AND
e. Clerical activities.
___ workflow following the abovementioned
activities shall be maintained at all times.
The prototype ___ ___ and the floor plan ___ for
constructing a COVID-19 testing laboratory shall be
used as references in constructing the testing
laboratory. (ANNEX D1 and ANNEX D2).
Controlled and adequate ___ with the prescribed ___
___ per hour shall be maintained for each specific
area.
Adequate ___ shall be provided in all areas.
LANGIT 2023
Features of Molecular Biology Laboratory Design:
10. Molecular Biology Laboratory Design
Prototype Floor Plan: COVID-19 Testing Laboratory
using RT-PCR
6.
Ante-Room:
a. specific area designed to maintain the pressure
differential between the laboratory and its adjacent
space
7.
Match: (a) Positive Pressure and (b) Negative Pressure
― Airflow inward
― Airflow outward
― No contaminated air will go out.
― No contaminated air will go in.
― Sample Preparation
― Reagent Preparation Room
― Isolation Rooms
― Amplification Room
11. Features of Molecular Biology Laboratory:
a. It consists of ___ ___, namely the (Enumerate):
i. ___ Room/Area
ii. ___ Room/Area
iii. ___ Room/Area
b. It operates in a ___ ___.
i. The sequence starts from: ___, ___ to ___.
ii. There must be no ___ ___. It must be
restricted to ___!
c. Its areas operate in a particular ___.
 Reagent Preparation Area → ___ Pressure
 Sample Preparation Area → ___ Pressure
 Amplification Area
→ ___ Pressure
d. Its areas consist of processes, some are in a
particular order yet some are not.
i. Reagent Preparation Area:
1. ___
2. ___
ii. Sample Preparation Area:
1. ___
2. ___
ii. Amplification Area:
1. ___
2. ___
Answer: BAABBABB
Page 10 of 15
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
13. Identify:
Molecular Biology Laboratory in FEU-NRMF
3.0 MOL BIO LABORATORY EQUIPMENT & INSTRUMENTS
Laboratory Equipment and Instruments:
 Autoclave
 Biological Safety Cabinet (BSC)
 Polymerase Chain Reaction Machine
 Spectrophotometer
 Vortex Mixer
 Centrifuge
 Automatic Pipettors
 Refrigerator
 pH Meter
12. Match:
a.
b.
c.
d.
e.
f.
g.
h.
―
―
―
―
―
―
―
―
Autoclave
Biological Safety Cabinet (BSC)
Polymerase Chain Reaction Machine
Spectrophotometer
Vortex Mixer
Centrifuge
Automatic Pipettors
pH Meter
is an instrument used to measure how much a
chemical substance absorbs light by measuring the
intensity of light as a beam of light passes through
sample solution.
very sensitive to changes in pH and hence, buffers
are used to stabilize the pH
It provides a physical method of sterilization by
killing bacteria, viruses, and even spores present in
the material put inside of the vessel using steam
under pressure (aka heat).
is a procedure that separates components of liquids
that have different weights. The device used for this
purpose is called a centrifuge, in which a centrifugal
force is applied that moves liquid components away
from the center.
are equipment used to deliver an accurately
measured volume of solution.
laboratory technique used to make multiple copies
of a segment of DNA
simple device used commonly in laboratories to mix
small vials of liquid
It is an engineering control intended to protect
laboratory workers, laboratory environment and
work materials from exposure to infectious or
biohazardous aerosols and splashes.
Answer: DHAFGCEB
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
Page 11 of 15
Vortex Mixer
Autoclave
RT-PCR
Automatic Pipettor
pH Meter
Centrifugre
Polymerase Chain Reaction (PCR) Machine
Spectrophotometer
Refrigerator
Biological Safety Cabinet
LANGIT 2023
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
(1) Autoclave:
LANGIT 2023
19. What makes the BSC efficient?
14. Autoclave:
a. It provides a ___ method of ___ by ___ bacteria,
viruses, and even spores present in the material put
inside of the vessel using ___ under ___ (aka ___).
b. What makes the autoclave efficient?
i. For efficiency, maintain ___ degrees
Celsius within ___ minutes at ___ psi
pressure.
c. Materials to be autoclaved depend on the ff factors:
i. ___
ii. ___
iii. ___
iv. ___
d. Materials such as:
i. ___ (shorter time and pressure)
ii. ___ (longer time and pressure)
20. HEPA Filters:
a. ___ (HEPA) Filters
b. They are constructed of pleated ___ glass and
arranged into ___ fibers.
c. It traps ___% of particles of ___ µm in diameter
(anything larger to ___ can be efficiently filtered by
this filter) since ___ and ___ are relatively smaller
they cannot be removed by this filtration.
15. Types of Autoclaves: Identify
21. Match:
a.
b.
c.
―
―
―
―
(2) Biological Safety Cabinet (BSC):
―
―
16. Biological Safety Cabinet (BSC):
a. It is an ___ control intended to protect laboratory
___, laboratory ___ and work ___ from exposure to
infectious or biohazardous aerosols and splashes.
b. Such aerosols and splashes may be generated while
___ materials containing infectious agents, such as
primary cultures, stocks and diagnostic specimens.
17. Identify:
―
―
―
―
Class I BSC
Class II BSC
Class III BSC
It is suitable for work in Biosafety Level 3 and 4
laboratories.
It is a ventilated cabinet, which provides personnel,
product and environmental protection.
It provides the highest level of personnel protection
It is commonly found in clinical and research
laboratories working with infectious agents in Risk
Groups 2, 3 and 4 (if positive-pressure suits are
used) or with tissue culture.
It is used for Risk Group 4 agents.
These are suitable for work with Risk Group 1
(RG1), Risk Group 2 (RG2), and Risk Group 3
(RG3) biological material.
simple Biological Safety Cabinet (BSC)
There are four types: A1, A2, B1, and B3. The main
differences between the types are the ratio of air
exhausted from the BSC to the air that is recirculated
within the BSC, and the type of exhaust system
present.
provides personnel and environmental protection but
not product production (as unsterilized room air is
drawn over the work surface)
It is the first designed
Answer: CBCBCAABAA
18. Factors that produce biohazard aerosols:
― Rapid hand movements
Working Mechanism:
― The working mechanism of these BSCs differs according to
their types. It has an open front with inward airflow for
personnel protection, downward HEPA filtered laminar
airflow over the work surface for product protection and
Page 12 of 15
BSMT 2, SEMESTER 2
―
―
―
―
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
HEPA filtered exhausted air for environmental protection. The
room air and recirculated air are HEPA filtered before flowing
downwards over the work area. These BSCs can be exhausted
into the containment zone or directly to the outside
atmosphere through a thimble or hard-ducted connection
depending on the type. The amount of air that recirculates or
exhausts depends on the type.
These BSCs should have an attached pass-through box that is
sterilizable and equipped with HEPA-filtered exhaust. Its
cabinet may be connected to a double-door autoclave used to
decontaminate all materials entering or exiting the cabinet.
The directional flow of air whisks aerosol particles that may
be generated on the work surface away from the laboratory
worker and are then discharged from the BSC through a
HEPA filter.
This type of cabinet is totally enclosed and is tested under
pressure to ensure that no particles can leak from it into the
room. Supply air is HEPA-filtered and exhaust air is
discharged into the atmosphere through two HEPA filters. The
operator access the work surface by means of heavy-duty
rubber gloves which form part of the cabinet. Several glove
boxes can be joined together to extend the work surface.
Airflow is maintained by a dedicated exhaust system exterior
to the cabinet, which keeps the cabinet interior under negative
pressure.
Room air is drawn in through the front opening at a minimum
velocity of 0.38 m/s. The front opening also allows the
operator’s arms to reach the work surface inside the cabinet
while he or she observes the work surface through a glass
window. The window can also be fully raised to provide
access to the work surface for cleaning or other purposes.
Answer: BCACA
22. Class 1 BSC:
a. It is the ___ designed and ___ Biological Safety
Cabinet (BSC) which provides ___ and ___
protection but not ___ production (as ___ room
air is drawn over the work ___).
b. These are suitable for work with Risk ___, Risk
Group ___, and Risk Group ___ biological
material.
23. Identify the letters.
Page 13 of 15
LANGIT 2023
24. Working Mechanism:
a. ___ air is drawn in through the ___ ___ at a
minimum velocity of ___ m/s. The front
opening also allows the ___’s ___ to reach the
___ ___ inside the cabinet while he or she
observes the work surface through a ___ ___.
The window can also be ___ raised to provide
access to the work surface for ___ or other
purposes.
b. The directional flow of air whisks ___ particles
that may be generated on the work surface ___
from the laboratory worker and are then ___
from the BSC through a ___ ___.
HEPA filter traps ___% of particles of ≥ ___ µm in
diameter. This enables the HEPA filter to effectively trap
all known infectious agents and ensure that only microbefree exhaust air is ___ from the cabinet and/or ___ in the
___ ___.
25. Class II BSC:
a. It is a ___ cabinet, which provides ___, ___ and
___ protection.
b. It is commonly found in ___ and ___
laboratories working with infectious agents in
Risk Groups ___, ___ and ___ (if ___-pressure
suits are used) or with ___ culture.
c. There are four types: ___, ___, ___, and ___.
The main differences between the types are the
___ of ___ ___ from the ___ to the ___ that is
___ ___ the BSC, and the ___ of ___ system
present.
i. About ___% of all biosafety cabinets
installed are Type ___ cabinets. There
is a limited need for Class II Type
___ biological safety cabinets.
ii. In addition, Class II Type ___
biological safety cabinets require very
specific ___ and operating ___ to
function correctly.
26. Identify the letters.
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
LANGIT 2023
31. Supply:
27. Working Mechanism:
a. The working mechanism of Class II BSCs
differs according to their ___. It has an ___ ___
with ___ airflow for ___ protection, ___ ___
___ ___ airflow over the ___ ___ for ____
protection and ___ filtered ___ air for ___
protection.
b. The ___ air and ___ air are ___ filtered before
flowing ___ over the ___ area.
c. Class II BSCs can be ___ into the ___ zone or
directly to the outside ___ through a ___ or ___
connection depending on the type.
d. The ___ of air that ___ or ___ depends on the
type.
28. Class III BSC:
a. It provides the ___ level of ___ protection and
is used for Risk Group ___ agents. It is suitable
for work in Biosafety Level ___ and ___
laboratories.
29. Identify the letters.
Table: Biological Safety Cabinet (BSC) Classes
Protection
Biosafety
Risk Group
Level
Agent
Laboratory
Class I
Class II Class
III
Class
Type
Table: Features of Class I, II, and III BSCs
Face
Airflow
Airflow
Exhaust
Velocity (recirculated) (exhausted) System
(m/s)
Class I
Class
IIA1
Class
IIA2
Class
IIB1
Class
IIB2
Class III
(3) Polymerase Chain Reaction Machine:
32. PCR Machine:
a. laboratory technique used to make ___ copies of a
segment of ___
33. Identify:
30. Working Mechanism:
a. This type of cabinet is ___ enclosed and is
tested under ___ to ensure that no particles can
leak from it into the room.
b. ___ air is ___-filtered and ___ air is ___ into
the atmosphere through ___ ___ filters.
c. The operator access the work surface by means
of heavy-duty rubber ___ which form part of
the cabinet. Several glove boxes can be joined
together to extend the work surface.
d. Airflow is maintained by a dedicated ___
system ___ to the cabinet, which keeps the
cabinet ___ under ___ pressure.
e. The Class III BSCs should have an attached
pass-through ___ that is ___ and equipped with
___-filtered exhaust. The Class III cabinet may
be connected to a ___ ___ used to ___ all
materials ___ or ___ the cabinet.
Page 14 of 15
BSMT 2, SEMESTER 2
1ST PRELIMINARY TERM
COURSE: MOLECULAR BIOLOGY & DIAGNOSTICS
(4) Spectrophotometer:
34. Spectrophotometer:
a. is an instrument used to measure how much a
chemical substance ___ ___ by measuring the ___
of light as a beam of light passes through ___ ___.
(5) Vortex Mixer:
35. Vortex Meter:
a. simple device used commonly in laboratories to ___
___ vials of ___
(6) Centrifuge:
36. Centrifugation:
a. is a procedure that ___ ___ of ___ that have
different ___.
37. Centrifuge:
a. The device used for this purpose is called a
centrifuge, in which a ___ force is applied that
moves liquid components away from the ___.
(7) Automatic Pipettors:
38. Pipettes:
a. are equipment used to ___ an accurately measured
___ of solution.
39. Micropipettes:
a. In a molecular biology laboratory, ___ are most
frequently used. Micropipettes measure volumes
that range from ___ μl (___ ml) to less than ___ μl.
(9) pH Meter:
40. pH Meter:
a. very sensitive to changes in ___ and hence, buffers
are used to ___ the pH
Page 15 of 15
LANGIT 2023
MOLECULAR BIOLOGY
DIAGNOSTIC LABORATORY
LABORATORY DESIGN,
EQUIPMENT AND
INSTRUMENTS
Molecular Biology Laboratory Design

LEARNING OUTCOMES :

1. To determine the Molecular Biology Laboratory design ( e.i. Covid-19 testing required by the
DOH)

2. To identify and know the functions of the instruments and equipment used in Molecular biology
lab

3. To determine the proper identification of the micropipette and to discuss the parts , functions
and principle of the different micropipette use in the mol bio lab.

4. To discuss the pH meter according to its parts, functions and principles.
ALLEN R. PENAFLORIDA,RMT,MSMT
Molecular Biology Laboratory Design
Molecular Biology Laboratory Design

According to DOH requirements :

According to DOH requirements :

https://doh.gov.ph/sites/default/files/health-update/ao2020-0014.pdf


PHYSICAL FACILITIES COVID-19 testing laboratories shall have adequate and appropriate areas to
safely, effectively and efficiently provide the services to clients.
https://doh.gov.ph/sites/default/files/healt
h-update/ao2020-0014.pdf

PHYSICAL FACILITIES COVID-19 testing
laboratories shall have adequate and
appropriate areas to safely, effectively
and efficiently provide the services to
clients.

2.Unidirectional workflow
following the above mentioned
activities shall be maintained at
all times.
1. There shall be a dedicated space for each of the following activities:
 a) Specimen reception;
 b) Virus inactivation and nucleic acid extraction (Pre-PCR);
 c) Reagent storage and handling;
 d) PCR; AND
 e) Clerical activities

Molecular Biology
Laboratory Design

According to DOH requirements :

https://doh.gov.ph/sites/default/files
/health-update/ao2020-0014.pdf

PHYSICAL FACILITIES COVID-19
testing laboratories shall have
adequate and appropriate areas to
safely, effectively and efficiently

3. The prototype floor plan and the
floor plan checklist for constructing
a COVID-19 testing laboratory shall
be used as references in
constructing the testing laboratory.
(ANNEX D1 and ANNEX D2) .
Molecular Biology
Laboratory Design
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES &
OTHER RESOURCES :
1-autoclave
Function
or purpose:
provides a physical method of
sterilization by killing bacteria,
viruses, and even spores present in
the material put inside of the vessel
using steam under pressure.
A list of autoclave part and their functions –
how to use an autoclave sterilizer
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :
1-autoclave

An autoclave is a pressurized chamber used for the
process of sterilization and disinfection by combining
three factors: time, pressure, and steam

Working Principle
•
Autoclaves use steam as their sterilization agent. The basic principle of an
autoclave is that all the items within the autoclave come in direct contact
with the steam for a particular period irrespective of the nature of the
material- whether it is liquid, plastic ware, or glassware.
•
The amount of time and the temperature depends on the type of material
being sterilized and the increase in temperature of the cycle allows for
shorter periods.

A list of autoclave part and their functions –
how to use an autoclave sterilizer
 An autoclave

•
•
A list of autoclave part and their functions –
how to use an autoclave sterilizer
•
Safety valve. is basically a fail-safe device that protects the
user from danger if all other electronic procedures fail to
function properly. Because of its essential role in safety, the
safety valve always needs to be inspected and tested
beforehand.
•
Cooling system. Before the waste-water coming from the
autoclave can enter the drain piping, it has to be cooled down
to avoid damage caused by the heat.
•
Vacuum system. Present in only certain types of autoclaves,
the vacuum system serves to replace all the air inside the
chamber with steam.
•
Steam generator. Also known as a boiler, a steam
generator’s purpose is to provide a source of steam for the
autoclave when there is no central source available.
Uses
Autoclaves are mostly used for the sterilization of
medical or laboratory equipment with the capacity of
sterilizing a large number of materials at once.
They are commonly used for the preparation of
culture media during laboratory applications.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :

FACILITIES & OTHER RESOURCES :
•
BIOLOGICAL SAFETY CABINET ( BSC)
•
BIOLOGICAL SAFETY CABINET ( BSC)
•
is an engineering control intended to
protect laboratory workers, laboratory
environment and work materials from
exposure to infectious or biohazardous
aerosols and splashes.
•
Class I BSC is the first designed and
simple Biological Safety Cabinet (BSC)
•
Such aerosols and splashes may be
generated while manipulating materials
containing infectious agents, such as
primary cultures, stocks and diagnostic
specimens.
HEPA filter traps 99.9% of particles of ≥ 0.3 µm in diameter.
This enables the HEPA filter to effectively trap all known
infectious agents and ensure that only microbe-free exhaust air
is discharged from the cabinet and/or recirculated in the work
surface.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :
•
BIOLOGICAL SAFETY CABINET ( BSC)

Class II Biological Safety Cabinet (BSC)

A Class II Biological Safety Cabinet (BSC) is a ventilated cabinet,
which provides personnel, product and environmental
protection. It is commonly found in clinical and research
laboratories working with infectious agents in Risk Groups 2, 3
and 4 (if positive-pressure suits are used) or with tissue
culture.

There are four types (A1, A2, B1, and B2 ) of Class II BSCs. The main
differences between the types are the ratio of air exhausted
from the BSC to the air that is recirculated within the BSC, and
the type of exhaust system present.

About 90% of all biosafety cabinets installed are Type A2
cabinets. There is a limited need for Class II Type B biological
safety cabinets. In addition, Class II Type B biological safety
cabinets require very specific installation and operating
conditions to function correctly.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
Working mechanism
FACILITIES & OTHER RESOURCES :
•
Sterile room with a laminar flow

FACILITIES & OTHER RESOURCES :
•
BIOLOGICAL SAFETY CABINET ( BSC)
Working mechanism
Class III Biological Safety Cabinet (BSC)
Class III BSC provides the highest level of
personnel protection and is used for Risk
Group 4 agents. It is suitable for work in
Biosafety Level 3 and 4 laboratories.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

Working mechanism
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :
Laminar Air Flow/ Laminar Hood
Laminar Hood is a closed device primarily for processes or instruments sensitive to microbial contamination.
Working Principle
•A Laminar Hood is made up of stainless steel, avoiding joints and corners to prevent the accumulation of
bacterial spores.
•This device creates a sterile environment with the flow of sterile air through a High-Efficiency Particulate Air
(HEPA) filter and shortwave ultraviolet germicidal lamp that sterilizes the workstation.
•Laminar Air Flow has to turn on 15 minutes before to ensure complete sterilization and the workstation
should be cleaned with ethanol before and after use.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES & OTHER RESOURCES :

Laminar Air Flow/ Laminar Hood
Uses
•Laminar Hood is commonly used to conduct processes that
are sensitive to contamination.
•It is used for experiments related to plant tissue culture and
for the experiments of genetic transformation.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :
•
Sterile room with a laminar flow
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
•Parts and functions :
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :

FACILITIES & OTHER RESOURCES :

Types of laminar flow cabinet

Types of laminar flow cabinet

Depending on the direction of movement of air, laminar flow cabinets are divided into two types:

Depending on the direction of movement of air, laminar flow cabinets are divided into two types:

1. Vertical laminar flow cabinet

2. Horizontal laminar flow cabinet
•
In the vertical flow cabinets, the air moves from the top of the cabinet directly towards the bottom of the cabinet.
•
•
A vertical airflow working bench does not require as much depth and floor space as a horizontal airflow hood which
makes it more manageable and decreases the chances of airflow obstruction or movement of contaminated air
downstream.
In the horizontal laminar flow cabinets, the surrounding air comes from behind the working bench, which is then
projected by the blower towards the HEPA filters.
•
The filtered air is then exhausted in a horizontal direction to the workplace environment.
•
One advantage of this cabinet is that airflow parallel to the workplace cleanses the environment with a constant
velocity.
•
The elluent air directly hits the operator, which might reduce the security level of this type of laminar flow cabinets.
•
The vertical laminar flow cabinet is also considered safer as it doesn’t blow the air directly towards the person
carrying out the experiments.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :
Procedure for running the LAMINAR FLOW CABINET
1.
Before running the laminar flow cabinet, the cabinet should be checked to ensure that nothing susceptible to UV rays is
present inside the cabinet.
2.
The glass shield of the hood is then closed, and the UV light is switched on. The UV light should be kept on for about 15
minutes to ensure the surface sterilization of the working bench.
3.
The UV light is then switched off, and a time period of around 10 minutes is spared before the airflow is switched on.
4.
About 5 minutes before the operation begins, the airflow is switched on.
5.
The glass shield is then opened, and the fluorescent light is also switched on during the operation.
6.
To ensure more protection, the working bench of the cabinet can be sterilized with other disinfectants like 70% alcohol.
7.
Once the work is completed, the airflow and florescent lamp both are closed and the glass shield is also closed.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER RESOURCES :

In conventional PCR

REAL TIME PCR MACHINE
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

FACILITIES & OTHER
RESOURCES :

REAL TIME PCR
MACHINE

VORTEX MIXER
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :
Micro Centrifuge
Micro centrifuge is a compact type of centrifuge ideal
for separating small liquid samples at high
speed (usually more than 6,000 rpm).
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
simple device used
commonly in laboratories to
mix small vials of liquid
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :
Mini Centrifuge
•For quick spin-downs of 5ml centrifuge tubes
•Includes adapters for 12x75mm tubes
•Conserves valuable bench space
•Starts and stops with closing/opening of the lid
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

PARTS OF THE Automatic
pipettors :

Automatic pipettors

Components of
micropipette

- PLUNGER

- TIP EJECTOR

-VOLUME DISPLAY

- TIP CONE
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

FACILITIES & OTHER
RESOURCES :

Automatic pipettors
/pipetting procedure

Automatic pipettors
/pipetting procedure

different methods:

different methods:

1. forward pipetting

2. Reverse pipetting
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

Automatic pipettors

Types of micropipette:

Micropipettes can be classified
depending upon:
1.Working Principle
2. Operating Mechanism
3. Number of Channels
4. Volume/Capacity
•Working Principle:
The plunger performs the following two functions1.Air Displacement Micropipette:
This type of micropipette works on the air displacement
principle.
1.Positive Displacement Micropipette:
In these micropipettes, the piston comes in direct contact of the
sample. The disposable tip in a positive displacement
micropipette is a microsyringe composed of a capillary and a
piston (movable inner part) which directly displaces the liquid.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

Automatic pipettors

Types of micropipette:

Micropipettes can be classified
depending upon:
1.Working Principle
2. Operating Mechanism
3. Number of Channels
•Operating Mechanism:
1.Mechanical Micropipette:
These micropipettes are operated manually based on a piston-shaft
spring mechanism.
2.Electronic Micropipette:
An electronic micropipette is mostly automated. The aspirating and
dispensing of liquid is performed by the one touch buttons instead
of manual plunger pressing and depressing. Electronic pipettes also
often enable the user to create custom programs on the device
allowing the pipettes to suit diverse application needs.
4. Volume/Capacity
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

FACILITIES & OTHER
RESOURCES :
Position
1
In this, the
micropipette is at
rest position.
Press fit a tip to the micropipette
without directly touching the tip.

Automatic pipettors

Automatic pipettors
Position
2

Using an Air Displacement
Micropipette:

Using an Air Displacement
Micropipette:
In this, the plunger is
depressed till the
first stop.
To aspirate the liquid in the tip,
press the plunger to the first stop.
Immerse the pipette tip vertically in
the liquid.
Release the plunger
Slowly release the plunger while the
tip is immersed. The liquid will be
aspirated into the pipette tip.
Liquid is filled in the tip as per
preset micropipette volume.
Depress the plunger
To dispense the liquid, place the tip
on the inner wall of the receiving
vessel at a steep angle.
Slowly press the plunger to the first
stop to dispense the liquid.
To empty the tip completely, press
the plunger to the second stop.
Wipe the tip on the inner wall while
taking the tip out of the vessel.
Position
3
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

Automatic pipettors

Types of micropipette:

Micropipettes can be classified
depending upon:
1.Working Principle
2. Operating Mechanism
3. Number of Channels
4. Volume/Capacity
•Number of Channels:
1.Single Channel Micropipette:
2.A single channel micropipette is the one which has a single
channel to aspirate or dispense the liquid.
3.Multi Channel Micropipette:
4.A multi channel micropipette has multiple channels to aspirate
or dispense the liquid. The commonly available multi channel
micropipette variants are the 8 channel, 12 channel and 16
channel. Multichannel micropipettes reduce the workload of a
single channel micropipette when working with large volumes of
samples.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

Automatic pipettors
Size and Range / Technical Specifications:
Micropipettes are available in different volumes ranging from
0.1 µl to 10,0000 µl. The commonly used variants of single
channel variable volume micropipettes are listed below along
with their permissible error limits as specified in the ISO 86552 standard.
They are sometimes referred to as P10, P20, P1000, P5000
pipettes based on the maximum volume that can be aspirated
/ dispensed using the pipette. For instance, a 0.5-10ul
micropipette may be commonly referred to as a P10 pipette.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

Automatic pipettors

How to choose the right micropipette?
1.Selecting the Type of Micropipette :
-air displacement micropipettes
- positive displacement micropipettes.
2. look at the sub types available based on criteria like the
volume to be handled, quality of tips, manual or electronic
pipettes, etc.
a. An adjustable micropipette or a fixed micropipette
b. You can either use a micropipette whose volume is already
fixed or one whose volume can be adjusted as per
requirement.
c. The volume to be handled
d. As a standard rule, it is better to choose the smallest
pipette capable of handling the required volume because
when the set volume is close to the minimum capacity of the
micropipette, the accuracy and precision of the readings
decrease.
1. Single channel or multichannel micropipettes:
2. This decision can be made based on the number of
samples or replicates you are working with. Single
channel micropipettes like the MICROLIT RBO Single
Channel (Fixed Volume and Variable Volume) are
used when the number of samples is less whereas a
large number of samples or well plates can be
easily handled by multichannel micropipettes like
the MICROLIT RBO Multichannel (8-channel and 12channel).
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

Automatic pipettors

Types of micropipette:

Micropipettes can be classified
depending upon:
1.Working Principle
2. Operating Mechanism
3. Number of Channels
4. Volume/Capacity
•Volume/Capacity:
1.Fixed Volume Micropipette:
In a fixed volume micropipette, the volume of liquid to be
aspirated or dispensed remains fixed. These micropipettes are
used when the same volume of liquid is to be dispensed multiple
times.
2.Variable Volume Micropipette:
This micropipette comes with a specific minimum and maximum
volume range. The volume of the liquid to be aspirated or
dispensed can be adjusted (within the instrument’s volume range)
depending upon the requirement of the user.
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

FACILITIES & OTHER
RESOURCES :

Automatic pipettors
DISPOSABLE TIPS
INSTRUMENTS AND EQUIPMENTS USED IN MOLBIO LAB

Automatic pipettors

How to choose the right micropipette?
1.Selecting the Type of Micropipette :
-air displacement micropipettes
- positive displacement micropipettes.
2. look at the sub types available based on criteria like the volume to be handled, quality of tips, manual or
electronic pipettes, etc.
3. Micropipette tips
4. Accuracy with specific tolerance
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB

FOR STORAGE OF SAMPLE

Stirling Upright ULT Freezer (-20°C to -86°C ) – SU780XLE
•
Full Size, 780 Liters of storage capacity
•
Highest storage to footprint ratio of any commercially ULT
freezer
•
-20°C to -86°C temperature setpoint range
•
Plugs into any AC outlet
•
Lowest energy consumption of any upright ULT
•
Store more than 58,000 vaccine vials
•
Integrated cold chain of custody tracking
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES & OTHER RESOURCES :
Using a pH Meter
• very sensitive to changes in pH and
hence, buffers are used to stabilize the
pH
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES & OTHER RESOURCES :
FACILITIES & OTHER RESOURCES :
pH Meter
pH Meter
measurement
system :
• The term pH is derived from “p,” the
mathematical symbol for negative
logarithm, and “H,” the chemical symbol
for Hydrogen.
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
consists of three parts: a pH
measuring electrode, a
reference electrode, and a
high input impedance meter.
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES & OTHER RESOURCES :
FACILITIES & OTHER RESOURCES :
pH Meter
measurement
system :
pH Meter
measurement
system :
consists of three parts: a pH
measuring electrode -a battery,
with a voltage that varies with
the pH of the measured solution.
consists of three parts: a
reference electrode
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES & OTHER RESOURCES :
FACILITIES & OTHER RESOURCES :
pH Meter
measurement
system :
pH Meter:
consists of three parts: a
high input impedance meter.
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
Temperature Compensation contained within the instrument,
because pH electrodes and
measurements are temperature
sensitive.
❑ Manual compensation
❑ Automatic temperature
compensation (ATC)
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
FACILITIES & OTHER RESOURCES :
FACILITIES & OTHER RESOURCES :
pH Meter:
pH Meter:
Buffer Solutions
Buffer Solutions
✓ that have constant pH values and the
ability to resist changes in that pH level.
✓ that have constant pH values and the
ability to resist changes in that pH level.
✓ They are used to calibrate the pH
measurement system (electrode and
meter)
FACILITITIES AND EQUIPMENTS USED IN MOLBIO LAB
✓ They are used to calibrate the pH
measurement system (electrode and
meter)
Molecular Biology Laboratory
References :
Laboratory biosafety manual, Third edition, World Health Organization (WHO), 2004.
A Guide to Biosafety & Biological Safety Cabinets. ESCD.
Richmond JY, McKinney RW. Primary containment for biohazards: selection, installation and
use of biological safety cabinets, 2nd ed. Washington, DC, U.S. Department of Health and
Human Services/Centers for Disease Control and Prevention/National Institutes of Health,
2000.
Health Canada. Laboratory biosafety manual, 2nd ed. Ottawa, Minister of Supply and Services
Canada, 1996.
Massachusetts Institute of Technology, Environment, Health & Safety Office, Biosafety Cabinets.
Molecular Biology Laboratory
Molecular Biology Laboratory
References :
References :
Pelt-Verkuil EV, van Belkum A, Hays JP. Principles and Technical Aspects of PCR Amplification.
Springer Netherlands Publishing, 2008. ISBN: 978-1-4020-6240-7 (Print) 978-1-4020-6241-4
(Online).
https://www.microlit.us/micropipette-product-guide/
Collecting, preserving and shipping specimens for the diagnosis of avian influenza A (H5N1) virus
infection. Guide for field operations. World Health Organization (Internet). Cited 2014 Sep.
Available from: http://www.who.int/csr/publications/surveillance/Annex7.pdf.
https://www.aphl.org/programs/newborn_screening/Documents/2016%20Molecular%20Works
hop/4.2%20-%20lab%20design%20QA%20QC%20considerations.pdf