(P) L1: Basic Lab Practices Flashcards
This protects you and your clothing from contamination, should not be worn outside the lab
lab coats
What type of footwear is appropriate for lab use?
closed shoes to protect the foot from punctures, and corrosive / infectious materials
When should gloves be changed?
when contaminated
these (2) equipment are worn when you are about to be in contact with hazardous aerosols and caustic chemicals
eyewear and masks
order of proper donning of PPE
gown, mask, goggles, and gloves
order of proper doffing of PPE
gloves, goggles, gown, mask
when should you wash your hands
before and after completion of work
T or F
Storing of food in the laboratory is fine as long as it is not in contact with the reagents
F
T or F
Applying cosmetics in the lab is fine as long as you wash your hands first
F
Where are sharps disposed of?
puncture-proof containers
T or F
Needles should be recapped from the syringes
F
This minimizes the creation of aerosols / splashes
closed containers
Where should infectious materials should be manipulated
biosafety cabinet
Which chemicals may be used to decontaminate work sufaces?
10% bleach or 70% ethyl alcohol
What steps (2) should be done before disposing infectious materials
placed in a biohazard bag or autoclaved
Where should incidents be reported?
appropriate personnel
Use this card to familiarize steps to do in a biological accident / emergency
- use safety showers for 15 minutes if exposed to any corrosive chemicals and bacterial culture splatters; use the eyewash if necessary
- wash with water and consult with the clinic if injured by contaminated sharp items (e.g. broken glassware, syringes, etc.)
- use paper towels in cleaning spillage of more than 1L of cell cultures (paper towels are to be autoclaved); decontaminate with 10% bleach or 70% ethyl alcohol
- evacuate if there is spillage of large volumes of hazardous chemicals
studies the structure and function of molecules and macromolecular systems associated with biological processes
molecular biology
Molecular biology focuses on the study of _____ and _______
nucleic acids and proteins
Molecular biology isolates and identifies _____ / ______ materials from different samples
genetic / protein
What are the typical methods employed in molecular biology?
PCR, gel electrophoresis, protein analysis, and sequencing techniques
T or F (Lab design)
Physical facilities of COVID testing shall have adequate and appropriate areas to provide service to the clients
T
T or F (Lab design)
Bidirectional workflow shall be maintained at all times
F (Unidirectional) str8 tayo d2
T or F. The airflow of a reagent prep room/area should be positive pressure.
T
What is the direction of positive pressure
outward
What is the direction of negative pressure?
inward
2 reasons for separate areas
- Introduction of unwanted nucleic acids may lead to contamination
- The sensitivity of molecular techniques makes them vulnerable to contamination
3 ways to control contamination
laboratory design, lab practices, and chemical and enzymatic controls
use this card to familiarize the potential sources of contamination
- cross contamination between specimens
- amplification of product contamination
- lab surfaces
- ventilation ducts
- reagents / supplies
- hair, skin, saliva, and clothes
a physical method of sterilization by killing bacteria, viruses, and spores
autoclave
How do autoclaves kill bacteria, viruses, and spores
it uses steam under pressure (heat)
What are the different levels of decontamination
- cleaning
- disinfecting
- sterilization
level of decontamination where bacteria is minimized
cleaning
level of decontamination where viable organisms are eliminated
disinfecting
level of decontamination where viable organisms and their spores are eliminated
sterilization
equipment that protects the workers, environment, and materials from infectious or biohazardous aerosols and splashes
Biological safety cabinet (BSC)
what are the three classes of BSC?
class I, II, and III
BSC characterized to have an open front with directional airflow that protects the personnel and environment using a HEPA filtered exhaust and operates under negative pressure
Class I
What is the biosafety level requirement of containment for class I BSC
1-3
What BSC class is characterized with an open front with directional airflow for personnel and environmental protection with HEPA-filtered laminar down flow for product protection
Class Ii
How many HEPA filter(s) is/are in a class I BSC
1
How many HEPA filter(s) is/are in a class II BSC
2 (HEPA exhaust and HEPA filter laminar downflow)
How many HEPA filter(s) is/are in a class III BSC
3
What are the different types of class II BSC
A1, A2, B1, B2
What is the most common type of Class II BSC
A2 (90%)
Why is type B Type II BSC not popular?
requires very specific installation and operating conditions
Which risk groups does Type II BSC work with (given that positive pressure suits are used)
2,3, and 4
where is the air discharged in type II BSC
rear plenum and into the space between supply and exhaust filters
how much air pass through the exhaust HEPA filters in class II BSC
30%
How much air is recirculated through the HEPA filter in class II BSC
70%
This class of BSC cabinet provides the highest level of protection
Class III
T or F
Class III biosafety cabinets has no holes at all and utilizes automated machines to handle infectious specimens
F there are holes sealed with gloves
Fume hood or BSC?
Filters and blowers are incorporated
BSC
Fume hood or BSC?
Protects the user only
Fume hood
Enumerate what the safety cabinet protects
personnel, product, environment (PPE)
Where does the vapors and aerosols go when using a fume hood
outside the building
T or F
Safety cabinets does not exhaust outside the building without decontamination
T
BSC types for personnel protection, microorganisms in risk group 1-3
Class I, II, and II
BSC type for personnel protection, microorganisms in risk group 4, glove box lab
class III
BSC type for personnel protection, microorganism in risk group 4, suit laboratory
Class I, Class II
BSC type for product protection
Class II, Class III (only if laminar flow included)
BSC type for volatile radionuclide / chemical protection, minute amounts
Class IIB1, Class IIA2 vented to the outside
BSC type for radionuclide / chemical protection
Class I, Class IIB2, Class III
T or F
Chemical fume hoods must be near the BSC
F (must not be near)
T or F
BSC must be near anything that created air movement for better ventilation
F (must not be near)
T or F
BSC must be far from the entrance, near the rear of the laboratory, away from traffic
T
T or F
BSCs are certified at initial installation and quarterly thereafter
F (annually thereafter)
T or F
BSCs must be certified after moving, replacement of filters, and maintenance
T
T or F
BSCs must be certified according to the manufacturer’s instructions
t
Use this card to familiarize yourself with the utilization of BSCs
- Magnahelic gauge should be checked regularly
- The airflow pattern should not be disrupted
- Disinfect the interior and materials
- Should run at least 5 minutes after the work volume is set up
- Open flames are to be avoided
- Decontamination
These are made from pleated borosilicate glass arranged in random fibers
HEPA filters
what does HEPA stand for
High efficiency particulate air
fill in the blanks
HEPA filters traps _____% of particles larger than _____ in diameter
99.97% of particles larger than 0.3 micrometer
This produces an amplicon that may be used for gel electrophoresis
conventional PCR
this checks the concentration and purity of sample
also measures how much a chemical substance absorbs light by measuring its intensity as a beam is passed through a sample solution
spectrophotometer
this mixes small vials of liquid
vortex mixer
this separates components of liquids that have different weights
centrifuge
this delivers an accurately measured volume of solution
automatic pipettors
what are the most frequently used type of pipettors
micropipettes
micropipette range measures are from ______ to less than _______
1000µl (1mL) to less than 1µl
T or F. The airflow of a sample preparation room/area should be positive pressure.
F (negative)
T or F. The airflow of a amplification room/area should be positive pressure.
F (negative)
In establishing a molecular testing in Clinical lab environments the workflow of rooms should be in what sequence?
Reagent prep room -> sample prep room -> ampli room
designed to maintain the pressure differential between the lab and its adjacent space; not all rooms have the same pressure.
ante room
T or F. The appropriate BSC for cell & tissue culture, propagation of virus is BSC 1.
F
This BSC is commonly found in clinical and research laboratories working with infectious agents or with tissue culture.
BSC 2
The class that is associated with BSL 1, 2, and 3 containments
BSC 2
air circulated and exhausted: A1 & A2
70% recirculated and 30% exhausted
air circulated and exhausted: B1
70% exhausted and 30% recirculated
air circulated and exhausted: B2
100% exhaust
Which of the following does not belong: BSC Class II air flow
A. Once room air enters, instead of directly proceeding to the work surface, it will first go to the open/front exhaust grill,
B. Then it will go down, then up, then some air
will be exhausted and some air will be recirculated
C. And then the air in the work surfaces will go
to the front or rear exhaust grill.
D. None of the above.
D
What BSC Class has a double HEPA filter exhaust, autoclave and dumptank for disposal.
3
What BSC class is suitable for work in Biosafety Level 3 and 4 laboratories.
3
The HEPA filter traps particles of what diameter?
0.3 micrometer in diameter
T or F. Anything larger to 0.3 can be efficiently filtered by HEPA filter such as vapor and gases.
F (gases and vapors r relatively smaller)
used to amplify a specific region of any DNA sample; the target is amplified
thermocycler
a type of PCR that uses gel electrophoresis and UV transilluminator
conventional
A microcentrifuge is designed for small tubes between ml?
0.2mL and 2mL
ideal ref temp
-20c and -40c
