Genetics Lab Exam 1

Front 1

Why is it necessary to clean the BSC with 10% bleach at the start of each exercise?

Back 1

Reduces risk of contaminating YPD-agar plate with non-yeast cells.

Front 2

Why is it necessary to clean the BSC with 10% bleach at the end of each exercise?

Back 2

After is to reduce the risk of contamination of future work under the hood.

Front 3

How does a Class II BSC provide both primary and secondary containment?

Back 3

By use of air curtain, plastic window, and HEPA filter.

Front 4

Why are tips disposed of in sharps waste rather than regular waste?

Back 4

To keep tips from poking through regular trash bags and scratching housekeeping staff during disposal.

Front 5

Volume range of P-20

Back 5

2-20 ul

Front 6

Volume range of P-200

Back 6

20-200 ul

Front 7

Volume range of P-1000

Back 7

200-1000 ul

Front 8

What types of micriobiological samples are spread by wands and loops?

Back 8

Wands spread cells in liquids. Loops spread solid masses of cells.

Front 9

MeSH

Back 9

Medical Subject Headings

Front 10

SOP's

Back 10

Standard Operating Procedures: Procedures for spills, emergencies, exposure incidents; methods for limiting exposure; in Chemical Hygiene Plan

Front 11

Risk Group 1

Back 11

Microorganism is unlikely to cause human or animal disease

Front 12

Risk group 2

Back 12

Agents are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available

Front 13

Risk group 3

Back 13

Agents are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available

Front 14

Risk group 4

Back 14

Agents are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available

Front 15

Purpose of containment

Back 15

To reduce exposure of laboratory workers, other persons, and the outside environments to potentially hazardous agents

Front 16

Primary containment

Back 16

the protection of personnel and the immediate laboratory environment from exposure

Front 17

Secondary containment

Back 17

the protection of the environment external to the laboratory from exposure

Front 18

Biosafety Level 1

Back 18

SOP: wash hands, restrict or limit access when working, no eating drinking smoking, safe sharps, decontaminate work surfaces
Lab type: Basic teaching, research
Lab practices: GMT
Safety equipment: open bench, PPE

Front 19

Biosafety Level 2

Back 19

SOP: Level 1+ PPE, mechanical pipetting
Lab type: Primary health services, diagnostics services, research
Lab practices: GMT plus protective clothing, biohazard sign posted
Safety equipment: open bench plus BSC

Front 20

Biosafety Level 3

Back 20

Lab type: special diagnostic services, research
Lab practices: Level 2+ special clothing, controlled access, directional air flow
Safety equipment: BSC and other devices for special activities

Front 21

Biosafety Level 4

Back 21

- Facility Design for secondary barriers: ante-room, negative air flow, autoclave within the lab
Lab type: dangerous pathogen units
Lab practices: Level 3+ air lock entry, shower exit, special waste disposal
Safety equipment: Class III BSC, positive pressure suits, double ended autoclave, filtered air

Front 22

When do you need to use the BSC?

Back 22

For work with infectious agents involving aerosols/splashes, large volumes, high concentrations

Front 23

Chemical Safety

Back 23

- Must have inventory of chemicals kept in lab
- An MSDS
- All chemicals must be clearly labeled, including hazards
- Chemicals should be segregated by hazard class
- Chemicals should be dated

Front 24

PPE

Back 24

- Gloves - latex, nitrile, other
- Face shield - full face coverage
- Goggles - wrap around, over glasses, contacts**
- Masks - respirator (fumes, particulates), paper (dust, particulates)
- Lab coats - long or short, sleeves
- Aprons - chemical resistant (wet, concentrated chemical)
- Hot/Cold gloves - rubber or cloth
- Eyewash station
- Safety shower

Front 25

When do you and don't you use primary containment?

Back 25

DO: infectious agents, tissue culture, bacteriology, rDNA
DON'T: organic solvents, dust

Front 26

Spill kits have

Back 26

Gloves, absorbent pads, absorbent worms, absorbent powder/pellets, broom, dustpan

Front 27

Waste disposal - Orange bags

Back 27

rDNA
- 90 minutes at 250 degrees F
- NO sharps
- Autoclave monthly QC testing
- Autoclave logbok
- Affix label to front after treatment

Front 28

rDNA =

Back 28

recombinant deoxyribonucleic acid

Front 29

Waste disposal - Red bag

Back 29

Human and Animal Tissue
- Tear/leak resistant
- Stored for no more than 7 days at room temp or 15 days under fridge at temp 35-45 degrees F
- Place in two red bags and packaged within rigid container prior to transport
- Incineration box dated and labeled with generator information
- Receptacles must be disinfected after red bag removal

Front 30

Centrifuges

Back 30

- Increase gravity
- Separate components by weight/density
- Used to move liquid to the bottom of the tube after vortexing, to pellet denser/heavier materials to separate them from the liquid medium

Front 31

What two events are required for stable transformation?

Back 31

Entry into cell (via LiAc) & double recombination between exogenous DNA and yeast chromosome

Front 32

What does PCR do...?

Back 32

Replicates specific segments of DNA

Front 33

What primers did we use for PCR?

Back 33

ADE2A and D to prime DNA synthesis/replication

Front 34

Steps of PCR

Back 34

Denature DNA
Anneal (primers attach)
Elongation (via Taq DNAP)

Front 35

How are cells lysed prior to PCR?

Back 35

Treatment with lyticase to degrade the cell wall and high temperatures to burst cell/nuclear membranes

Front 36

How is lyticase inactivated?

Back 36

heated to 95 degrees before PCR

Front 37

Tris-HCl in PCR

Back 37

maintains optimum pH for PCR

Front 38

KCl in PCR

Back 38

provides physiological concentrations of salt (high salt PCR buffer -> shorter PCR product; low salt PCR buffer -> amplifying long PCR product)

Front 39

MgCl2 in PCR

Back 39

provides Mg++ ions needed by Taq DNA polymerase

Front 40

dNTP in PCR

Back 40

building blocks from which the DNA polymerase synthesizes a new DNA strand

Front 41

Primers ADE2A and D in PCR

Back 41

flank and amplify the gene of interest of the yeast genome in both WT and T

Front 42

non-transformed s288c a cells in PCR

Back 42

serve as a template for the primers in colony PCR; serve as a control when comparing amplicons of transformed cells