OHS026
Safe Work Procedure
Faculty/Division
School/ Divisional Unit
SCHOOL OF MEDICAL SCIENCES
MEDICINE
Document number
Initial Issue date
20-4-10
Current version
Current Version
Issue date
Next review date
24/4/2013
SOMS GBD SWP
The Writing Safe Work Procedures Guideline (OHS027) should be consulted to assist in the completion of
this form.
Safe Work Procedure Title and basic
description
Title: Labeling synthetic RNA probe (T7/SP6) with Digoxigenin
Description: Use Roche DIG RNA labelling Kit (SP6/T7) to label newly synthesised RNA strand with DIGUTP in T7/SP6 reverse transcriptase reaction.
Associated risk assessment title and location: Use PCR machine; potential electric shock or burn from
hot-plate.
Describe the activity or process
- Add PCR product cDNA to reaction mix containing NTP, Buffer, RNasin in RNase free PCR tube.
- Vortex and spin briefly.
- Incubate at 37°C for 2hrs in PCR machine.
- Remove from PCR machine, cool on ice.
- Read OD.
- Check labelling efficiency by “dot test” on Nitrocellulose paper.
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Page 1 of 2
Safe Work Procedure
Date Effective: 01/01/2007
Uncontrolled document when printed
Current Version: 1.2, 15/08/2007
List all resources required including plant,
chemicals, personal protective clothing and
equipment, etc
 Lab gown, closed shoes, gloves;
 PCR tubes, Bio-Rad absorbance cuvettes.
List potential hazards and risk controls
including specific precautions required
 DIG kit reagents can cause irritation to eyes and skin by contact or inhalation. Wear gloves and
gown and safety glasses when handling the kit. Must read MSDS FOR KITS before the start of the
experiment.
 Use Eye wash and shower station if contact occurs.
 All work is RNase sensitive. Must wear gloves and gown.
 When loading the PCR machine, be aware that the base can be hot. Do not touch it.
List emergency shutdown instructions
 In the event of inhalation or ingestion the first aid officer should be consulted immediately.

First Aid Officer: Renee Szokolai ext 58497
List clean up and waste disposal requirements
 Wash and autoclave all tips and glassware.
 Wear gloves all the time to protect samples from degradation.
 Plastic tubes and plates should be disposed in correct biological waste bags.
List legislation, standards and codes of practice
used in the development of the SWP
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: 2006 Safety in laboratories Part 3: Microbiological aspects and containment facilities
AS/NZS 1336:1997 Recommended Practices for Occupational Eye Protection
AS/NZS 2161.1:2000 Occupational Protective Gloves – Selection, Use and Maintenance
Supervisory approval, training, and review
Supervisor: Prof E Burcher
Signature:
Plant custodian:
Signature
List competency required – qualifications, certificates, licencing, training - eg course or instruction:
COMMON SENSE
SWP review date: 24/4/2013
Responsibility for SWP review:Fei Shang
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Page 2 of 2
Safe Work Procedure
Date Effective: 01/01/2007
Uncontrolled document when printed
Current Version: 1.2, 15/08/2007