M1: INTRODUCTION TO MOLECULAR BIOLOGY LABORATORY

Question 1

An ideal laboratory consists of three completely independent rooms or areas

Answer 1

reagent preparation room
sample preparation room
PCR room

Question 2

This part of the laboratory is identified as clean rooms

Answer 2

Pre-amplification rooms

Question 3

This part of the laboratory is identified as dirty rooms

Answer 3

Post-amplification rooms

Question 4

This room should be under positive air pressure

Answer 4

Pre amplification room

Question 5

What does positive air pressure do ?

Answer 5

impedes the entrance of any airborne contaminant into the pre-amplification room when the door is opened

Question 6

What does negative air pressure do ?

Answer 6

impedes anything from coming out of this laboratory and contaminating the surrounding environment

Question 7

This is referred to as the PCR room

Answer 7

Post amplification room

Question 8

Pre-amplification rooms include

Answer 8

reagent preparation room

sample preparation room

Question 9

Describe the unidirectional workflow of a molecular biology laboratory

Answer 9

reagent prep -> sample prep -> amplification -> post amplification

Question 10

The construction of this into each laboratory is an inexpensive way of creating differential pressure for the laboratory

Answer 10

anteroom

Question 11

Safety precautions for anterooms

Answer 11

door leading into the anteroom from the outside and the door to the inside should never be opened at the same time

adding sticky mats at the entrance of each anteroom laboratory

Question 12

This pertains to containment principles, technologies and practices that are implemented to prevent the unintentional exposure to biological material or their accidental release

Answer 12

biosafety

Question 13

Pertains to the protection, control and accountability for biological agents and toxins within facilities in order to prevent their loss, theft, misuse, diversion, unauthorized access, or intentional unauthorized release

Answer 13

biosecurity

Question 14

Safety concerns on inactivation

Answer 14

virulence and amount of pathogens
sense and mobility limitations of wearing full PPE
user hygiene and health status
spills and aerosol-generating procedures
harsh chemicals
exposure to extreme heat and cold
glassware

Question 15

Safety concerns on Nucleic acid extraction

Answer 15

check if procedures have prior inactivation
irritating chemicals
spills and aerosol generating procedures
harsh chemicals
exposure to extreme heat and cold
glassware

Question 16

Safety concerns on PCR and other nucleic acid tests (Pathogen is inactivated)

Answer 16

Disposal is still in biohazard bags for environmental concerns
Cleanliness maintained for proper detection
Hazards from equipment (heat, entrapment, moving parts)

Question 17

Safety concerns on Post-PCR (Pathogen is inactivated)

Answer 17

contamination from liquids with amplified nucleic acid

hazards from electricity, UV, chemicals, heat, sharps

Question 18

This type of waste includes all utensils, containers, tissue paper, etc that have been in contact with biohazardous material

Answer 18

biohazard waste

Question 19

This type of waste is usually segregated in strong yellow coloured bags

Answer 19

biohazard waste

Question 20

This type of waste is usually collected in yellow bench-top bins

Answer 20

sharps

Question 21

document that gives detailed information about a material and about any hazards associated with the material

Answer 21

Material Safety Data Sheet (MSDS)

Question 22

written guideline stating the policies, procedures, and responsibilities that serve to protect laboratory personnel from the health hazards associated with the hazardous chemicals

Answer 22

Chemical Hygiene Plan (CHP)

Question 23

Why should you never use a metal spatula when working with peroxides ?

Answer 23

because metals will decompose explosively with peroxides

Question 24

used to track chemicals from their time of purchase to when they are used and discarded

Answer 24

Chemical tracking system

Question 25

Patient specimens from suspected or confirmed cases should be transported as

Answer 25

UN3373, “Biological. Substance, Category B”

Question 26

Viral cultures or isolates should be transported as

Answer 26

Category A, UN2814, “infectious substance, affecting humans”

Question 27

What is a HEPA filter

Answer 27

High Efficiency Particulate Air Filter

Question 28

This biosafety cabinet is also known as open front BSC

Answer 28

BSC 1

Question 29

This BSC does not protect the sample/ product from possible contamination

Answer 29

BSC 1

Question 30

This BSC protects the worker from potentially infectious aerosols; Also capable of protecting the sample from possible contamination

Answer 30

BSC 2

Question 31

What is the principle of BSC 2 ?

Answer 31

Air passes through HEPA filter BEFORE flowing over the materials within [Protects the specimen from contamination]

Air to be exhausted first passes through HEPA filter [Protects the worker and the environment from potentially infectious aerosols]

Question 32

What type of BSC 2 has 70% of air is recirculated to the working area; 30% of air is exhausted ?

Answer 32

BSC Class IIA

Question 33

What type of BSC 2 has 30% of air is recirculated to the working area; 70% of air is exhausted ?

Answer 33

BSC Class IIB1

Question 34

What type of BSC 2 has no recirculation of air; Total exhaust of air through an exhaust HEPA filter ?

Answer 34

BSC Cass IIB2

Question 35

What type of BSC is completely enclosed and are equipped with glove ports; Infectious material within is handled with rubber gloves that are attached and sealed

Answer 35

BSC 3

Question 36

Trained laboratory staff or certified contractors should perform tests or check on Adjustable and fixed-volume automatic pipettors

Answer 36

Check for volumetric accuracy and reproducibility, and recalibrate as necessary before placing in service initially and at specific defined intervals, at least once every 6 months.

Pipettor inaccuracy is one of the most common sources of laboratory error.

Question 37

Trained laboratory staff or certified contractors should perform tests or check on Thermometers

Answer 37

Test against a standardized thermometric device annually

Question 38

Trained laboratory staff or certified contractors should perform tests or check on Refrigerators and freezers

Answer 38

Establish tolerance limits for temperatures. The refrigerator’s temperature tolerance limits
must reflect the most stringent needs of all reagents, supplies, and specimens stored within it.

Maintain daily records of temperatures and appropriate documentation of corrective action for out-of-range temperatures.

Question 39

Trained laboratory staff or certified contractors should perform tests or check on Incubators and water bath

Answer 39

Establish tolerance limits for temperatures, carbon dioxide level, and humidity as applicable.

Maintain daily record of temperatures and appropriate documentation of corrective action for out-of-range
temperatures.

Question 40

Trained laboratory staff or certified contractors should perform tests or check on Centrifuges

Answer 40

Measure operating speeds periodically with a tachometer. Document the readings.

Maintain daily record of temperatures for refrigerated centrifuges. Verify performance of centrifuge timers by comparing to a known standard every 6 months.

Question 41

Trained laboratory staff or certified contractors should perform tests or check on Autoclaves

Answer 41

Verify content processing using heat sensitive tape with each autoclave batch.

Verify effective sterilization with an appropriate biological indicator weekly.

Perform autoclave maintenance annually or as per manufacturer, including a pressure check and calibration of temperature device.

Check autoclave mechanical timing device periodically.

Maintain records of autoclave operation and maintenance in the equipment log.

Question 42

Trained laboratory staff or certified contractors should perform tests or check on Analytical balances

Answer 42

Check accuracy with standard weights and class at a predetermined frequency. Document the results with an evaluation of their capacity.

Service and calibrate periodically using qualified personnel.

Maintain records of service and calibrations.

Place the analytical balance in a stable location so that vibration does not affect the readings.

Question 43

Trained laboratory staff or certified contractors should perform tests or check on Biosafety cabinets/laminar air flow hoods

Answer 43

Verify air intake grills are not obstructed.

Certify cabinets/hoods annually by a trained service technician, certified maintenance department or company.

Check daily for air flow as instructed by manufacturer and document the results to verify the effectiveness of the hood’s personnel and environmental protective
functions.

Clean the ultraviolet lamp, if used, weekly with 70% alcohol.

Document daily and weekly cleaning.

Question 44

Trained laboratory staff or certified contractors should perform tests or check on Generators

Answer 44

Maintain and keep readily accessible records to verify that the backup generator system is in place and
operational.

Follow SOPs that detail generator maintenance. Procedure should list the frequency and procedural steps of maintenance and testing.

Supporting logs should document monthly checks on critical generator components, including fluid levels (oil, coolant, and fuel), belts, battery, testing startup, and operation.

Question 45

Trained laboratory staff or certified contractors should perform tests or check on Out of service equipment

Answer 45

Any equipment that is out of service for any reason should be clearly identified and removed from the laboratory