1 | P a g e

RISK ASSESSMENT OF HAZARD IN GENETIC ENGINEERING LABORATORIES

By

NIK ABDUL HAFIIDZ BIN NIK ABDUL MALEK

2014688106

SAIFUL ANWAR BIN NAZURALLAH

2014626794

AHMAD AHKMAL BIN MASRUDIN

2014631198

MOHAMMAD ARSHAD BIN RASHID

2014683386

This report is prepared for

MADAM RAFEQAH BINTI RASLAN

SEPTEMBER 2015

2 | P a g e

TABLE OF CONTENTS

Chapter 1 - Hazard Identification

1.1 Hazard and Biohazard

1.2 Identify Hazards 1.3 Hazard And Biohazard In Experiment 1.3.1 Laboratory 1 Gene in Bottle 1.3.2 Laboratory 2 Bacterial Transformation 1.3.3 Laboratory 3 Restriction Endonuclease Digestion And Gel Electrophoresis Of Dna

1.3.4 Laboratory 4 Crime Scene Investigator PCR 1.3.5 Laboratory 5 Enzyme Linked Immunosorbent Assay (ELISA) Chapter 2 - The Biological Risk Assessment

Chapter 3 - Biohazard Equipment 3.1 Enzyme Linked Immunosorbent Assay (ELISA) 3.2 Genes in the bottle 3.3 Crime Scene Investigator PCR 3.4 Bacterial Transformation 3.5 Restriction endonuclease digestion and gel electrophoresis of DNA

Chapter 4 - Emergency Response Procedure/ Report/ Accident

Chapter 5 - Recommendation For Improvement In Fkk Laboratory

3 | P a g e

Chapter 1

Hazard Identification

In Malaysia, laboratories can be found in universities, company’s research and development

and government institute. The laboratory environment can be a hazardous place to work.

Laboratory workers such as students are exposed to numerous potential hazards including

chemical, biological, physical and radioactive hazards, as well as musculoskeletal stresses.

Laboratory safety is governed by numerous local, state and federal regulations. The reduction

of hazards improves the safety and quality of life for human and for the environment, these

hazards can be minimized through researches and improve safety procedure (Maaza, 2008).

Therefore performance of risk assessment of hazardous chemical and biohazard is an

important process to organize the management of hazards of these chemicals and biohazard.

This study was conducted to assess the risk from genetic engineering laboratory course in

chemical and bioprocess engineering program.

1.1 Hazard and Biohazard

Hazard is defined anything for example condition, behaviour, situation, practice that has the

potential to cause harm, including disease, death, injury, environmental, property and

equipment damage. A hazard can be a thing or a situation. (Western Sydney University, 2015)

According to Western Sydney University (2015), hazard Identification is the process of

examining each work area and work task for the purpose of identifying all the hazards which

are “inherent in the job”. Work areas in this report specifically on laboratories however it also

apply to other work area for example office areas, agricultural stores and transport,

maintenance and grounds. This process is about finding what could cause harm in work task

or area specifically on laboratories.

Biological hazards, also known as biohazards, are organic substances that pose a

threat to the health of humans and other living organisms. Generally speaking, biological

hazards include pathogenic micro-organisms, viruses, toxins (from biological sources),

spores, fungi and bio-active substances. Biological hazards can also be considered to include

biological vectors or transmitters of disease. Outside the health arena, biological hazards

include substances that cause social and economic disruption, property damage and

environmental degradation, such as insect plagues or infestations. Worldwide, it is estimated

4 | P a g e

that around 320 000 workers die each year from communicable diseases caused by work-

related exposures to biological hazards (Safe Work Australia, 2011).

1.2 Identify Hazards

Identify hazards is the first step in risk management step in the workplace. A hazard is anything

including work practices or work practice or procedures that have the potential to harm the

health and safety of a person. The student is responsible for identifying hazards which may

be present in the workplace. Laboratory safety requires knowledge of laboratory procedures,

equipment, and reagents, as well as constant watchfulness for danger. One careless student

can cause accident to other student. Hazard is classified as below:

Electrical hazard are sources of danger from electrical equipment that can cause

electrical shock Example of Electrical equipment placement of cords, wet areas, hot plates.

Potential impact electrocution and fire;

Chemical hazard are sources of danger from exposure to laboratory chemicals,

including immediate and long-term effects on the health of workers. Chemicals are identified

by using MSDS and summarized or fill in a table. Potential Impact are lung irritant, flammable

and more.

Physical hazard are sources of danger in the environment, including, housekeeping

accidents, and falls. Example of Physical water spills on floor, hot, cold, appropriate storage,

pressure, gas cylinders; Potential Impact are personal injury, falling, slipping.

Biohazard are sources of danger from living (“bio”) specimens, including blood and

other body fluids, microbiology specimens, and cultures. Example of biohazard is infectious

organisms, samples likely to contain infectious organisms; Potential Impact is health risks;

5 | P a g e

Figure 1: The risk assessment procedure (Western Sydney University, 2015)

Lab specimens sometimes contain disease-causing microorganisms, called pathogens.

Exposure of lab personnel to pathogens is likely to vary from one medical practice to another.

A small rural family practice clinic will have a much different patient population with different

health problems than will a specialty practice in a large metropolitan area. Nonetheless,

general principles of hygiene and safety should be followed in all POLs to decrease the risk of

disease transmission. Biohazards potentially infective biospecimens encountered in clinical

laboratories include the following:

• blood

• body fluids

• body tissue biopsies

• urine

• exudates (pus, mucus, sputum)

• bacterial smears

• bacterial cultures

1.3 Hazard And Biohazard In Experiment

6 | P a g e

During genetic engineering laboratories course in semester 4, 5 experiment was conducted.

Only one experiment involving the use of microorganism and other experiment mostly involves

the use of enzyme and DNA. All the experiments are classified in risk group 1 and biosafety

level 1. All the experiment are low risk of hazard. There are 4 potential hazard during

experiment which is biohazard, chemical hazard, physical hazard and electrical hazard. The

possible hazard and experiment information as follow below.

1.3.1 Laboratory 1 Gene in Bottle

In this experiment there is no biohazard involve. However, the swab use to obtain human

cheek cell should be clean and avoid any bacteria and other hazardous material enter the

mouth. The chemical use in this experiment cool isopropanol and soap which is not hazardous

to heath but are highly Flammability. Electrical equipment use is centrifuge which should be

handle carefully. Physical hazard may occur according to statement above. the aim of this

experiment is to isolate gene.

Figure 2 : gene isolated after centrifuge.

1.3.2 Laboratory 2 Bacterial Transformation

The experiment involve the use of microbacteria E. Coli which is classify under biosafety level

1 and risk group 1. E. Coli is a microorganism that is unlikely to cause human or animal disease

but the high concentration of microorganism that enter the body may cause diarrhea. In this

7 | P a g e

experiment, the E.coli cell was transform by adding new foreign gene which is pGlo to observe

its bacterial transformation. The newly transformed bacteria should be keep safe and avoid in

contact with human body. The aim of experiment is to develop an understanding of the

biological process of bacterial transformation by the pGLO plasmid DNA. Physical and

chemical hazard may occur according to statement above.

Figure 3: transformed bacteria (E. Coli) is covered to avoid contaiminantion to sample and

avoid exposure.

1.3.3 Laboratory 3 Restriction Endonuclease Digestion And Gel Electrophoresis Of

Dna

The aim of this experiment is to separate and sort a large group of DNA molecules by size,

determine the size of each molecule separated and develop an understanding of the role of

restriction enzymes and agarose gel electrophoresis to cut and size DNA. The possible

biohazard in this experiment is the enzyme use in this experiment. There are probability that

when enzyme enters into human body it may cause sickness. The major hazard in this

experiment is during gel electrophoresis, the buffer used which contain electric current should

be keep safe and do not touch the buffer liquid . it may cause electrical shock. Physical hazard

may occur according to statement above.

8 | P a g e

Figure4: Agarose gel before (left) and after (right) electrophoresis procedure

1.3.4 Laboratory 4 Crime Scene Investigator PCR

In this experiment, the procedure quite familiar to the restriction endonuclease digestion and

gel electrophoresis of dna lab only the sample where change for the electrophoresis. The

major hazard involve in this experiment is during electrophoresis. The dna sample of the

experiment is consider non-biohazard. No microbial use in this experiment. Physical, chemical

and electrical hazard should be avoided.

Figure 5: The PCR tube containing DNA sample, mastermix and primers.

1.3.5 Laboratory 5 Enzyme Linked Immunosorbent Assay (ELISA)

9 | P a g e

The aim of this experiment is study the basic ELISA procedure to measure the concentration

of antibodies or antigens in solutions. In this experiment, the potential biohazard is sample

taken from student which is human saliva. It may contain some bacteria. When exposed to

another student it may cause low risk of infection depends on that type of virus or infection

that student carries.

(A) (B)

Figure 6: Result of ELISA’s test shows patients with antigen (left) and patients without antigen

(right).

---------------------------------------------------------------------------------------------------------------------------

Chapter 2

The Biological Risk Assessment

Risk assessment is an important responsibility for directors and principal investigators of

microbiological and biomedical laboratories. Institutional biosafety committees (IBC), animal

care and use committees, biological safety professionals, and laboratory animal veterinarians

share in this responsibility. Risk assessment is a process used to identify the hazardous

characteristics of a known infectious or potentially infectious agent or material, the activities

that can result in a person’s exposure to an agent, the likelihood that such exposure will cause

an infection. The information identified by risk assessment will provide a guide for the selection

of appropriate biosafety levels and microbiological practices, safety equipment, and facility

safeguards that can prevent harm.

Our laboratory manager and laboratory assistant use risk assessment to alert their

staffs to the hazards of working with infectious agents and to the need for developing

proficiency in the use of selected safe practices and containment equipment. This procedure

10 | P a g e

used in our UiTM Bioprocess Laboratory. Successful control of hazards in the laboratory also

protects persons not directly associated with the laboratory, such as other occupants of the

same building, and the public. This follow the Occupational Safety & Health Act 1994,

USECHH 2000 Regulation.

The Lecturer or laboratory assistant must consult with a Laboratory manager to ensure

that the laboratory is in compliance with established guidelines and regulations. When

performing a risk assessment, it is advisable to take a conservative approach if there is

incomplete information available. Factors to consider when evaluating risk include the

following:

Pathogenicity

Route of transmission

Agent stability

Infectious dose

Concentration

Origin

Availability of data from animal studies

Availability of an effective prophylaxis or therapeutic intervention

Medical surveillance

Experience and skill level of at risk personnel

In this assignment I need to categorize the risk group, biosafety level and safety containment

level based on the experiment that we have done in genetic laboratory experiment. We need

to ensure that all the experiment follow the guideline.

1) RISK GROUPS

In many countries, including the United States, biological agents are categorized in Risk

Groups (RG) based on their relative risk. Depending on the country or organization, this

classification system might take the following factors into consideration:

Pathogenicity of the organism

Mode of transmission and host range

Availability of effective preventive measures (e.g., vaccines)

Availability of effective treatment (e.g., antibiotics)

Other factors

11 | P a g e

It is important to understand that biological agents are classified in a graded fashion such that

the level of hazard associated with RG1 being the lowest and RG4 being the highest. EHS

Biosafety follows the NIH Guidelines categorization of Risk Groups as follows:

RG1 – Are not associated with disease in healthy adult humans or animals

RG2 – Are associated with disease which is rarely serious and for which preventative

or therapeutics is often available

RG3 – Are associated with serious or lethal human disease for which preventative or

therapeutics may be available

RG4 – Are associated with lethal human disease for which preventative or therapeutics

are not readily available

RISK GROUP 1 AGENTS: REQUIRING CONTAINMENT LEVEL 1

Risk Group 1 (low individual and community risk)

This group includes those microorganisms, bacteria, fungi, viruses and parasites, which are

unlikely to cause disease in healthy workers or animals.

RISK GROUP 2 AGENTS: REQUIRING CONTAINMENT LEVEL 2

Risk Group 2 (moderate individual risk, limited community risk)

A pathogen that can cause human or animal disease but under normal circumstances, is

unlikely to be a serious hazard to healthy laboratory workers, the community, livestock, or the

environment. Laboratory exposures rarely cause infection leading to serious disease; effective

treatment and preventive measures are available and the risk of spread is limited.

RISK GROUP 3 AGENTS: REQUIRING CONTAINMENT LEVEL 3

Risk Group 3 (high individual risk, low community risk)

A pathogen that usually causes serious human or animal disease, or which can result in

serious economic consequences but does not ordinarily spread by casual contact from one

individual to another, or that can be treated by antimicrobial or antiparasitic agents.

RISK GROUP 4 AGENTS: REQUIRING CONTAINMENT LEVEL 4

Risk Group 4 (high individual risk, high community risk)

A pathogen that usually produces very serious human animal disease, often untreatable, and

may be readily transmitted from one individual to another, or from animal to human or vice-

versa directly or indirectly, or casual contact.

12 | P a g e

2) BIOSAFETY LEVELS

In contrast to Risk Groups, Biosafety Levels (BSL) prescribe procedures and levels of

containment for the particular microorganism or material (including Research Involving

Recombinant or Synthetic Nucleic Acid Molecules). Similar to Risk Groups, BSL are graded

from 1 – 4. Detailed descriptions of containment practices and biosafety levels can be found

in our laboratory guidelines.

The majority of experiments done at UiTM Bioprocess Laboratory involves Biosafety Level

1 and 2 (BSL-1) & (BSL-2) practices. BSL-2 containment and practice is suitable for work with

agents that are infectious to humans or animals where exposure may result in limited to

moderate disease. The routes of exposure to these agents are typically through cuts and

breaks in the skin, ingestion, and splashes to the mucous membranes (eyes, nose, and

mouth). These agents or materials include:

1. Microorganisms (e.g., RG2 or higher)

2. Human blood, blood components, fluids, unfixed organs, tissues and cell lines (primary

and established)

3. Non-Human Primate Derived Materials (including established cell lines)

4. Biotoxins (with and LD 50 of less than 100 micrograms per kilogram of body weight in

vertebrates) requiring BSL-2 containment

5. Research Involving Recombinant or Synthetic Nucleic Acid Molecules

13 | P a g e

Table 1: The Biosafety Levels

THE RISK CATEGORIZE BY EACH EXPERIMENT.

EXPERIMENT 1 – GENES IN A BOTTLE

This is the abstract of the experiment. DNA stands for deoxyribonucleic acid, a genetic

material vital to a living thing. They can be collected from any bodily parts, the most common

ones includes blood, hair and saliva in a human. Physically, DNA can only be seen under the

microscope. But if enough DNA is available from lysed cells combines and intertwines with

each other, it becomes visible to the eyes as whitish strands. The aim of this experiment was

to perform DNA extraction and precipitation and to observe a collection of DNA strands. This

was done by collecting cheek cells from inside the mouth to extract the DNA as they divide

very often as new cells replace them. After collecting the cheek cells, it was mixed and

incubated with a series of chemicals and apparatus such as a lysis buffer and protease to

extract the DNA. The experiment was deemed successful as a collection of DNA was

successfully extracted and precipitated. Afterwards, the DNA collection was collected into a

vial for observation and keepsake.

In this experiment, we don’t use any harmful microorganism but the harm may come

when we need to remove the protein using high temperature. From the method that we used

14 | P a g e

in this experiment, we can conclude that it can be put under risk group 1, containment level 1

and biosafety level 1.

EXPERIMENT 2 – BACTERIAL TRANSFORMATION

This is the abstract of the experiment. Transformation is basically the process in which a

bacterium takes up an exogenous DNA and expresses its genes, resulting in changes to its

traits. One of the commonly used genes to demonstrate bacterial transformation is a gene that

codes for Green Flourescent Protein (GFP). When exposed to long ultraviolet (UV) wave light,

it emits bright green light that is visible in bacteria transformed by plasmid that contain the

genes encoding GFP. In this experiment, bacteria were transformed using transformation

solutions and heat shock procedures. The bacteria used in this experiment is a commonly

used bacteria used for genetic transformation is the Escherichia coli, or E. coli. And the

plasmid used to transform the bacteria is the pGLO plasmid, which encodes a gene for GFP

and a gene for resistance to the antibiotic ampicillin. These characteristics of the plasmid

provide a viable screening and selection process for transformed bacteria. Adhering to the

objectives of the experiment, an understanding of the biological process of bacterial

transformation by the pGLO plasmid DNA was developed and the acquired phenotypic trait of

GFP exhibited by transformed bacterial was observed.

In this experiment, we deal with Escherichia coli, or E. coli which is the microorganism

that classified under risk group 2. In order to deal with the bacteria, we must follow the

containment level 2, thus this experiment also can be categorize under biosafety level 2

EXPERIMENT 3 – RESTRICTION ENDONUCLEASE DIGESTION AND GEL

ELECTROPHORESIS OF DNA

This is the abstract of the experiment. Restriction enzymes are like scissors for DNA. They

are biomolecules that restrict (cut), DNA at specific sites. Agarose gel electrophoresis is a

powerful separation method frequently used to analyse DNA fragments generated by

restriction enzymes. Electrophoresis refers to the migration of a charged molecule through a

restrictive matrix, or gel, drawn by an electrical force. This experiment aimed to separate and

sort a large group of DNA molecules by size and determine it, as well as to develop an

understanding of the role of restriction enzymes and agarose gel electrophoresis to cut and

size DNA. Agarose gel electrophoresis was successfully used to separate and sort a large

group of DNA molecules by size. This is apparent through the different bands showed by the

agarose gel after the electrophoresis. The size of each molecule separated was determined

using two methods: direct gel examination and standard curve of the semi-log graph.

15 | P a g e

In this experiment, we use restriction enzyme as the main microorganism. The restriction

enzyme not harmful if only at our skin but it may be harmful if it is inserted inside the human

body. This experiment also deal with high voltage and high temperature equipment. From what

we have done in this experiment, it can be categorize under risk group 2 that lead to

containment level 2 and also can be put under biosafety level 2.

EXPERIMENT 4 – THE ISOLATION OF GENETIC MATERIAL BY PCR

This is the abstract of the experiment. This experiment aims to demonstrate and understand

the techniques of PCR, analyze and evaluate its results and also to determine its sensitivity.

This was done in the need of DNA fingerprinting or DNA profiling for a crime scene

investigation. The sample from the crime scene as well as the DNA for a group of suspects

was put under the procedures of PCR. Following the procedure, the amplifying process was

easily achieved.

In this experiment, there are no biological harmful effect because all the sample was already

prepared by lab assistant. The risk that may involve is when we want to transfer the genetic

from a tube to another and also we need to deal with high temperature and high voltage

equipment. Therefore we can categorize them under risk group 2 that lead to containment

level 2 and also can be put under biosafety level 2.

EXPERIMENT 5 – ELISA

This is the abstract of the experiment. The Enzyme – linked immunosorbent assay (ELISA)

uses antibody to detect the presence of a disease agent such as viruses, bacteria and

parasites in the blood or other body fluid. This experiment demonstrates the application of

ELISA in real situation. The enzyme, HRP is essential to present the positive result of the

assay where it oxidizes the colorless substrates into a blue solution. The blue color indicates

that there are antigens present in the serum and the antibodies have successfully bound onto

them. This experiment was fairly successful since all the wells gave out different result for

different samples, but accordingly to the theory. Moreover, the results that was taken into

consideration was only the color changes that occurred within 5 minutes. Therefore, the results

are deemed acceptable.

In this experiment, the sample used is unknown but it is under risk group 2 microorganism.

These experiment also can detect the harmful microorganism such as virus SARS or even

HIV. That virus can be categorize under risk group 4. In our level although the sample are

unknown or maybe in form of GMO, the risk group involve is in group 2. In order to handle it,

we must follow the biosafety level 2 or 3 and the containment level 2.

16 | P a g e

Chapter 3

Biohazard Equipment

While handling experiment in the laboratory, many safety precaution needs to follow by person

to make sure each experiment finish with success. Before uses every equipment in the

laboratory, each person needs to read the sign on that particular equipment. These show how

to handling without any problem happens.

Figure 7: Sign on Laminar Flow Hood

General rules need by everyone that enter the laboratory is everyone in the lab is

responsible for their own safety. Before starting any work, personnel should be familiar with

procedures and equipment used to handle any experiments. Basic needs by personal safety

are lab coats, gloves, goggles, shoes and masks need apply during in the laboratory.

17 | P a g e

Figure 8: Sign for personnel safety

Besides personnel responsibilities, housekeeping and decontamination needs after

each experiment finish. Work areas must be kept clean and free from any chemicals. Throw

all the chemical that already uses into chemical waste tank. Clean up equipment to avoid any

possibility of exposing of chemical into next personnel. Furthermore, do not block any

emergency equipment to make sure during any accident happen all the emergency equipment

can be use and this will reduce personnel exposure to chemical.

Figure 9: Chemical waste

18 | P a g e

Figure 10: Fire extinguisher

Figure 11: Fire extinguisher sign

19 | P a g e

Figure 12: Eyewashes Figure 13: Regular check card

Every laboratory also need smoke alarm to detect fire before huge fire happen and

damage chemical. Explosion in laboratory will damage the entire chemical in the storage and

this will increase exposure of poison smoke into person. Exposures of chemical are very

dangerous because it will widely expose through the air and move about 2km and more around

the accident place.

Figure 14: Smoke detector

20 | P a g e

When any accidents happen in the laboratory, every person must take immediately

precautions to help themselves. Emergency maps must shown at every building to make sure

each person move out from that building in safety condition to the assembly point. Follow the

exit sign that shown on the laboratory door to the assembly point.

Figure 15: Sign of fire procedure Figure 16: Emergency sign

Figure 17: Exit sign

21 | P a g e

Table 2: Biohazard equipment

3.1 Enzyme Linked Immunosorbent Assay (ELISA)

ELISA uses antibody to detect the presence of a disease agent such as viruses, bacteria and

parasites in the blood or other body fluid. ELISA have been used in pregnancy test, disease

detection in people, plant and animals, detecting illegal drug use, test for indoor air quality and

test to determine whether the food is labelled accurately. In general, an antigen is any

molecule that can provoke the immune system to respond. This immune response is specific

to a type of antigen (Panigrahi, B. K., Abraham, A., & Das, S., 2010). There are two options of

ELISA detection which are direct detection and indirect detection.

Goggles Lab Coat

NAME OF EXPERIMENT

CHEMICAL USES/ MICROORGANISM

USES

LAMINAR FLOW HOOD

MASK LAB COAT

GOGGLES GLOVES

Elisa Phosphate buffer

Genes in the bottle

Lysis buffer

Isopropanol

The isolation of genetic material by PCR

Orange G loading dye

TAE buffer

Bacterial transformation

Escherichia coli

Calcium Chloride

Restriction endonuclease digestion and gel electrophoresis of DNA

Buffer solution

EcoRI

Pstl

HindIII

22 | P a g e

Gloves Masks

Figure 18: Safety Equipment use in lab

3.2 Genes in the bottle

Protein synthesis involves two main steps: transcription and translation. The DNA plays an

important role as the first molecule that is needed in the process. The letters of the nucleotides

encoded in the DNA is copied by enzymes, basically forming 3 types of RNA; the messenger

RNA (mRNA) the transfer RNA (tRNA) and the ribosomal RNA (rRNA) (Brown, 2010). These

experiment shows DNA extraction and precipitation and to observe a collection of DNA

strands. DNA that been collecting from human body was check cells to use in these

experiment. To break open the cells to release the DNA, lysis buffer contains detergent, will

been used. Then, to make the DNA visible cold alcohol is used. Using isopropanol, the DNA

is able to get out of the solution in which it is in, and form clumps to differentiate itself. To

handle both of these chemicals, mask, goggle, gloves and lab coat must been used while

handling during the experiment.

3.3 The isolation of genetic material by PCR

It is a polymerase chain reaction (PCR) technique that determines the alleles present at

different short tandem repeats (STR) loci within a genome in order to use DNA information to

identify individuals (Andreas, Nicole and Dimitri, 2004). From this experiments, the objective

to demonstrate and understand the techniques of PCR, to evaluate the results of PCR and to

determine the sensitivity of PCR.

3.4 Bacterial transformation

Genetic transformation has given rise to many successful findings in terms of medicinal,

agricultural and bioremediation aspects. For example, agriculture sector have been modified

to have better resistance towards pests and other damaging factors, as well as to improve its

23 | P a g e

growth. Bacteria have also been transformed to assist in bioremediation processes, in terms

of pollutant absorption and elimination. E.coli will be used for genetic transformation. Like all

bacteria, E. coli does not have any nuclear membrane or a nucleus, thus indicating that all of

the genes required for its survival are only found in a single chromosome (Karp and Pruitt,

1996). To handle E.coli, mask, gloves, goggles, lab coat and laminar flow hood was been used

to prevent E.coli from death and affect human body from get diarrhea.

Figure 19: Laminar Flow Hood used to avoid exposure to the microorganism

3.5 Restriction endonuclease digestion and gel electrophoresis of DNA

Restriction enzymes are like scissors for DNA. They are biomolecules that restrict (cut) DNA

parts. Agarose gel electrophoresis is a powerful separation method frequently used to analyze

DNA fragments generated by restriction enzymes (Mozayani, 2011). This experiment aimed

to separate and sort a large group of DNA molecules by size and determine it, as well as to

develop an understanding of the role of restriction enzymes and agarose gel electrophoresis

to cut and size DNA. All the restriction enzyme uses was transferred into tubes. Then, DNA

that through process of cutting fragment by restriction enzyme will move to electrophoresis

process.

24 | P a g e

Chapter 4

Emergency Response Procedure/ Report/ Accident

From our interview with the secretary of safety and health committee of FKK, Miss Hajatun

Rabani binti Ahmad Razif. Till today, there are only one major accident which are the

laboratory ceiling dropped but it did not cause any injuries. Most common accident occurs in

the laboratory are broken beakers which mostly caused by mishandling. There are no serious

injury due to accidents in the laboratory that are reported until today.

Provided below is the form used to report any accident occurs in the laboratory, the

first person that need to be informed first are the lab assistant to measure the severity of the

accident for further action needs to be taken. From the form, we can see that any accident are

treated very seriously in spite of their severity. This is to comply with the FKK safety and policy

which is to continually improved compliance with OSH legislations and effective

implementation of FKK UiTM OSHMS by achieving and maintaining certification to OHSAS

18001:2007 and MS1722:2011.

Also provided below is the standard emergency procedure when fire occurs in the

laboratory, this procedure must be shown and followed by all personnel that works in the

laboratory to prevent chaos and reduce risk of injury in case of fire occurs.

Chapter 5

Recommendation For Improvement In Fkk Laboratory

Up till today, there are no accident that have caused serious injuries or illness in FKK

laboratories. This proves that the preventive and proactive action taken by the management

is successful. However, there is always room for improvement. There are few suggestion that

can be taken into consideration by the management in order to increase the safety of the

laboratory.

Place a printed policy of statement of environment, health and safety from the faculty

Organize a talk on safety which involves management, employees, faculty, staff and

students.

Make it a requirement for new employees and students to go through safety orientation

to expose them to the equipment they will use and explain the hazard of the chemical

that will be use in the future.

Involve students in Safety and Health committee.

25 | P a g e

Give incentives to employees and students whom exhibit excellent performance in

safety.

Ensure students to read safety manual of the equipment that they will be using along

with the lab manual.

Conduct announced lab inspection to ensure all equipment are in the best condition at

all time.

Students must not be allowed to work alone in the lab without approval from the staff.

Every experiment must be monitored until finish unless they have failsafe.

Lecturer must explain the hazard of the chemical that will be used and PPE that are

required before conducting experiments.

References

A & A SCIENTIFIC RESOURCES SDN BHD, 2011. Malaysia’s 1st University Affiliated

Environmental Laboratory. Retrieved on 27 September 2015 from

http://www.environment.com.my/about/

Andreas Manz, Nicole Pamme and Dimitri Lossifidis (2004), Bioanalytical Chemistry; Imperial

College Press,

Anonymous 2014. 40 steps to improve lab safety. Retrieved September 27, 2015, from

http://safety.nmsu.edu/programs/lab_safety/steps_to_lab_saf.htm

Brown, T. (2010). Gene cloning and DNA analysis: an introduction: Wiley-Blackwell.

FKK, 2012. Ohsasform/Accident Incident Investigation Report retrieved on September 27,

2015, from http://fkk.uitm.edu.my/v1/Ohsasform/Accident Incident Investigation

Report (1).pdf

Karp, G., & Pruitt, N. L. (1996). Cell and molecular biology: concepts and experiments: John

Wiley & Sons New York.

Maaza Yousuf Abdulah Mohamed, 2008. Risk Assessment Of Hazardous Chemicals A Case

Study In Chemistry Research Laboratories Of USM. University Sains Malaysia

Mozayani, A. (2011). The forensic laboratory handbook procedures and practice: Springer

Science+ Business Media.

Safe Work Australia, 2011. National Hazard Exposure Worker Surveillance – Exposure to

biological hazards and the provision of controls against biological hazards in Australian

26 | P a g e

workplaces retrieved on 26 september 2015 from

http://www.safeworkaustralia.gov.au/sites/SWA/about/Publications/Documents/571/N

HEWS_BiologicalMaterials.pdf

Western Sydney University, 2015. Hazard Identification, Risk Assessment and Control

Procedure. Retrieved on 26 September 2015 from

http://www.uws.edu.au/__data/assets/pdf_file/0020/12917/12917_Hazard_Identificati

on,_Risk_Assessment_and_control_Procedure.pdf

27 | P a g e

Appendix