MOL BIO LAB L5 (Midterms- Lab Set-up)

Question 1

The laboratory should be compatible with ________ to prevent contamination

Answer 1

mechanical barrier

Question 2

This is the most common issue in the molecular laboratory

Answer 2

contamination

Question 3

What area does the sample preparation room belong to?

Answer 3

Area 1

Question 4

What area does the Amplification room belong to?

Answer 4

Area 3

Question 5

What area does the Reagent Preparation room belong to?

Answer 5

Area 2

Question 6

An amplification room/area where the gel electrophoresis room is located

Answer 6

Analysis of PCR Products

Question 7

serves as a clean bench area

Answer 7

Dead airbox with UV light

Question 8

An amplification room/area where the PCR room is located

Answer 8

PCR amplification

Question 9

Room/ area in the molecular biology lab where biosafety cabinet (optional) is located

Answer 9

Nucleic acid Extraction Room/ Area

Question 10

The only room/area in the molecular biology lab where there is positive air pressure

Answer 10

Reagent preparation room/area

Question 11

Air pressure in Sample prep and Post amplification rooms/areas

Answer 11

Negative air pressure

Question 12

Single entrance, reagents used for amplification should not be exposed to other areas

Answer 12

Reagent prep

Question 13

Specimens should not be exposed to post-amplification work areas

Answer 13

Specimen prep

Question 14

The size of each area should consider ________ for equipment and ________ needed for preparation

Answer 14

space; bench space

Question 15

Alternative to Spatial Separation

Answer 15

• Class II biological safety cabinet
• Dedicated areas for each work phase
• Unidirectional
• Automated specimen processing station/closed tube amplification and detection system

Question 15

State the other lab design considerations (at least three)

Answer 15

• Temperature and humidity requirements
• Exhaust ventilation
• Water quality
• Electric outlet
• Back-up power system
• Eyewash
• Ergonomic assessment

Question 16

This is always included when checking for contamiantion

Answer 16

blank (no template) control

Question 16

What is to be avoided when pipetting?

Answer 16

Production of aerosols

Question 16

The PCR workstation is cleaned at every?

Answer 16

Start and end of the workday/run

Question 17

The PCR workstation is cleaned using?

Answer 17

Uv light, 70% ethanol, fresh 10% sodium hypochlorite, DNA Away

Question 18

This induces thymidine dimers and other modifications that render nucleic acid inactive as a template for amplification

Answer 18

Ultra-violet light irradiation

Question 18

Work stations should be cleaned with __________, followed by removal with _________

Answer 18

10% sodium hypochlorite solution (bleach; ethanol and water

Question 19

Substitution of uracil (dUTP) for thymine (dTTP) during PCR amplification

Answer 19

Enzymatic inactivation with uracil-N-glycosylase

Question 20

Contaminating PCR amplicons are degraded leaving only genomic DNA avaialble for PCR

Answer 20

New PCR sample reactions pre-treated with Uracil-N-glycosylase (UNG)

Question 21

Wipe test (swab test) is performed?

Answer 21

Monthly

Question 22

This is used to detect, localize, and remove contamination and identify the source of the contamination

Answer 22

Wipe test (swab test)

Question 22

Proficiency testing is performed?

Answer 22

Twice a year

Question 23

Used for assessment of the Competence in Testing

Answer 23

Proficiency Testing

Question 24

If specimens are not commercially available alternative proficiency testing program has to be?

Answer 24

established (specimen exchange etc.)

Question 25

Addresses emergencies from an all-hazards approach. This establishes policy-and guidance ensuring that critical functions continue and that personnel and resources are relocated to an alternate facility in case of emergencies.

Answer 25

Continuity of Operations Plan (COOP Plan)

Question 26

Uses of molecular laboratory in the field of clinical diagnostics

Answer 26

1. Health 2. Medicine 3. Forensics 4. Pharmaceutical Industry 5. Biological warfare 6. Drug Discovery 7. PCR based Technology 8. Fluorescence in situ Hybridization (FISH) 9. Biochip 10. Peptide Nuicleic acid (PNA) 11.Proteomic Technology : 12. Electrochemical Detection of DNA

Question 27

Potential sources of contamination are?

Answer 27

- Cross contamination between specimens - amplification product contamination - laboratory surfaces - ventilation ducts - reagents/supplies and - hair, skin, saliva, and clothes of laboratory personnel.

Question 28

Contamination may cause?

Answer 28

- Incorrect results - Require extensive-cleanup - Loss of creditability - Impact on financial and performance

Question 29

Contamination can be controlled using proper?

Answer 29

- Laboratory-design - Laboratory practices - Chemical and enzymatic controls

Question 30

Nucleic acids used be stored/ used in what temp?

Answer 30

4-8°C or -15 to -25°C

Question 31

A fundamental skill in molecular biology, as it ensures that reagents and samples are measured accurately and reliably

Answer 31

Precise pipetting

Question 32

These pipettes are used for handling small volumes, typically in the microliter range

Answer 32

Micropipette

Question 33

They are essential for PCR, DNA quantification, and sample transfers.

Answer 33

Micropipette

Question 34

These are used for transferring larger-volumes, typically in the milliliter range. Often used for media preparation, cell culture, and larger-scale molecular biology experiments.

Answer 34

Serological pipettes

Question 34

Multichannel pipettes have multiple channels, usually _______, that allow for simultaneous pipetting of multiple samples. They are commonly used in high-throughput applications.

Answer 34

8 or 12

Question 35

Two commonly used techniques in pipetting

Answer 35

Forward and reverse

Question 36

The more traditional-and widely used technique

Answer 36

Forward pipetting

Question 37

Effective when working with viscous or volatile samples

Answer 37

Reverse pipetting

Question 37

This technique reduces the risk of over-pipetting and ensures the intended volume is accurately delivered

Answer 37

Reverse pipetting

Question 38

Suitable for general sample transfers and dilutions

Answer 38

Forward pipetting

Question 38

This technique is commonly used when precise volume measurements are required, such as in quantitative assays where even minor deviations can affect results

Answer 38

Forward pipetting

Question 39

This technique is ideal for transferring sample with unique characteristics, like highly viscous substances or volatile solutions

Answer 39

Reverse pipetting

Question 40

Often used in the preparation of high-concentration reagents and for DNA/RNA extractions

Answer 40

Reverse pipetting

Question 41

Essential to maintain the accuracy and reproducibility of experiments.

Answer 41

pH measurement

Question 41

Measures the hydrogen ion concentration in a solution

Answer 41

pH

Question 42

Formula to calculate dilutions

Answer 42

(Concentration stock) X (volume of stock used) = (Concentration final) X (final volume)

Question 43

Monitor all steps of analytical procedure

Answer 43

Quality Control Plan

Question 44

Measurement to monitor and record specific activities as part of the quality

Answer 44

Quality Indicator - Turn around time - % of failed runs - Population medium - Calibrator parameters - Graph to identify trend or shift - Monitor frequency and acceptable range