Medicine
School of Medical Sciences, Oncology research and muscle development units (CGM)
26.6.09
1.0
26.6.09
26.6.12
For additional information refer to the OHS Risk Assessment and Control Procedure, the OHS Risk Rating Procedure and the Hierarchy of Risk Controls.
SiRNA transfection Risks
Step 1: Identify the activity
SiRNA transfection involves the uptake of small interfering RNA, by cells in culture, using a transfection reagent such as lipofectamine 2000.
Procesure:
1. Cells are seeded in a 6 well plate 24h before transfection (1x10 5 cells/well).
2. 24 hr after seeding, transfection is conducted as follows.
3. Incubate SiRNA (80pmol/well) in serum free medium for 5 min
4. Incubate lipofectaime (5ul /well) with serum free medium for 5 min
5. Mix SiRNA and Lipofectamine in a single tube and incubate for further 15 min.
6. Add 500 µl of tranfection reagent to each well, swirl plate gently and place in humidified incubator.
7. A repeat transfection is carried out 24h later in exactly the same way and cells are harvested 24 h after second transfection.
8. A mismatch RNA construct that does not interact with any known mammalian genes was used as the transfection control (use same procedure as above).
*SFM-serum free medium
Location of Activity: Tissue culture room (lab 501)
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Risk Assessment and Control Form
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Current Version: 2.6, 16/07/2008

Step 2: Identify who may be at risk by the activity
The user and any other tissue culture workers present.
Steps 3 to 7: Identify the hazards, risks, and rate the risks
1. An activity may be divided into tasks. For each task identify the hazards and associated risks.
2. List existing risk controls and determine a risk rating using the UNSW Risk Rating Procedure.
3. Additional risk controls may be required to achieve an acceptable level of risk. Re-rate the risk if additional risk controls used.
Tasks
Hazards
(Step 3)
Associated risks
(Step 4)
Existing risk controls
Risk rating with existing controls *
(Step 5)
C L R
Use of Neuroblastoma cell lines
Potential contact with cancer cell lines
Low risk of developing a tumour
Wear latex gloves Minor
(2)
D L
Additional risk controls required
(Step 6)
(Apply the hierarchy of risk controls)
Not required
Risk Rating with additional controls *
(Step 7)
C L R
Use of DMEM culture medium
Potential biohazard
Inactivate all waste with bleach prior to disposal
Minor
(2)
D L Not Required from the UNSW Risk Rating Procedure * C = consequence L = likelihood R = risk rating
Step 8 Documentation and supervisor approval
(signature) Completed by: Melissa Desouza
Step 9: Implement the additional risk controls identified
Authorised by: Peter Gunning (signature)
Indicate briefly what additional risk controls from Step 6 above were implemented, when and by whom.
Risk control: Date: Implemented by:
Date: Jun09
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Risk Assessment and Control Form
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Date Effective: 01/01/2007
Current Version: 2.6, 16/07/2008

Risk control:
Risk control:
Risk control:
Date:
Date:
Date:
Implemented by:
Implemented by:
Implemented by:
Risk control: Date: Implemented by:
Step 10: Monitor and review the risk controls
It is important to monitor risk controls and review risk assessments regularly. Review is required when there is a change in the process, relevant legal changes, and where a cause for concern has arisen. Reviews could be scheduled on an annual basis. If the risk assessment has substantially changed a new risk assessment is warranted.
Review date: Reviewed by: Authorised by:
Review date:
Review date:
Review date:
Reviewed by:
Reviewed by:
Reviewed by:
Authorised by:
Authorised by:
Authorised by:
Review date: Reviewed by: Authorised by:
Documentation
It is a requirement that legal and advisory documentation that supports this risk assessment be listed. Such documentation includes Acts, Regulations, Australian Standards and Codes of Practice, where applicable.
NSW OHS Act 2000
NSW OHS Regulation 2001
Code of Practice for the Labelling of Workplace Substances
AS/NZS 2243.2:2006. Safety in laboratories. Part 2: Chemical aspects
AS/NZS 2243.3-2002. Safety in laboratories. Part 3: Microbiological aspects and containment facilities.
AS/NZS 2161.1:2000 Occupational Protective Gloves – Selection, Use and Maintenance
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Risk Assessment and Control Form
Uncontrolled document when printed
Date Effective: 01/01/2007
Current Version: 2.6, 16/07/2008

OHS697
What you need to do
1. Consider what can go wrong that can hurt someone
2. Determine what the most likely outcome would be - Consequences
3. Determine how likely those consequences are - Likelihood
4. Calculate the risk rating
5. Required action
CONSEQUENCES:
Severe
Major
Moderate
Minor
Insignificant
LIKELIHOOD:
Almost certain
Likely
Possible
Unlikely
Rare
LIKELIHOOD
Almost certain
A
Likely
B
Possible
C
Unlikely
D
Rare
E
Risk level
Very high
High
Medium
Low
How severely could someone be hurt death or permanent disability to one or more persons hospital admission required medical treatment required first aid required injuries not requiring first aid
How likely are those consequences? expected to occur in most circumstances will probably occur in most circumstances could occur at some time is not likely to occur in normal circumstances may occur only in exceptional circumstances
Insignificant
1
M
M
L
L
L
Minor
2
H
M
M
L
L
CONSEQUENCES
Moderate
3
H
H
H
M
M
Major
4
VH
H
H
M
M
Severe
5
VH
VH
VH
H
M
Required action
Act immediately:
The proposed task or process activity must not proceed. Steps must be taken to lower the risk level to as low as reasonably practicable using the hierarchy of risk controls.
Act today:
The proposed activity can only proceed, provided that:
(i) the risk level has been reduced to as low as reasonably practicable using the hierarchy of risk controls;
(ii) the risk controls must include those identified in legislation, Australian Standards, Codes of
Practice etc.
(iii) the risk assessment has been reviewed and approved by the Supervisor and
(iv) a Safe Working Procedure or Safe Work Method has been prepared.
(v) The supervisor must review and document the effectiveness of the implemented risk controls.
Act this week:
The proposed task or process can proceed, provided that:
(i) the risk level has been reduced to as low as reasonably practicable using the hierarchy of risk controls;
(ii) the risk assessment has been reviewed and approved by the Supervisor and
(iii) a Safe Working Procedure or Safe Work Method has been prepared.
Act this month:
Managed by local documented routine procedures which must include application of the hierarchy of controls.
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UNSW Concise OHS Risk Rating Table Effective date: 01/01/2007
Uncontrolled document when printed Current Version: 2.6,16/07/2008