STANDARD OPERATING PROCEDURE
Procedure:
School/Department:
SOP prepared by:
DNA or RNA extraction using spin columns
School of Biological Sciences
Angela Nikolic and Nick Coleman (SMB) edit M Joseph
Section 1 - Personal Protective Equipment
1. Lab coat or lab gown (buttoned up)
2. Nitrile or latex gloves
3. Proper enclosed footwear
4. Safety glasses (at stages where ‘binding buffer’ is used)
Section 2 – Potential Hazards + Safety precautions
1. Eye damage if corrosive chaotropic salts contact eyes. Wear safety glasses.
2. Poisoning if chaotropic salts ingested.
3. Skin damage if chaotropic salts contact bare skin. Wear PPE.
4. Environmental damage if chaotropic salts disposed of incorrectly (eg. down
sink). Don’t put spin column waste down the sink – it should be disposed of by
autoclaving (if recombinant) or as hazardous chemical waste (via room 225).
5. Environmental damage if recombinant DNA is disposed of incorrectly
6. Injury due to incorrect use of centrifuge. Do not use the centrifuge unless an
experienced user, preferably the equipment custodian, has given you a
thorough demonstration of how to use it correctly and safely.
7. Fire or electrocution due to centrifuge malfunction. Check the rotor for signs of
wear and tear before use, and retire rotors that are excessively scratched or
damaged. (check with the equipment custodian). Do not exceed speed rating
of the rotor. Ensure rotor is firmly attached to the spindle before starting machine.
Section 3 – Procedure
1. Follow procedure as outlined in the manual provided in your spin column kit (eg.
Qiaquick kit). Briefly, this procedure is outlined below.
2. Mix DNA with binding buffer, add to column, then spin in microcentrifuge.
3. Discard eluate into dedicated waste container
4. Add wash buffer to column, let sit 5 minutes, then spin in microcentrifuge.
5. Discard eluate into dedicated waste container
6. Dry top part of column eg. by opening lid, sitting on Kimwipe at ~50 C for 10 min.
7. Add elution buffer to column, let sit 5 minutes, then spin in microcentrifuge.
8. Retain DNA-containing eluate for further work, discard plastic column parts into
autoclave waste.
Section 4 – Disposal / Spills / Incidents
1. Don’t put spin column waste down the sink – dispose by autoclaving (if
recombinant DNA present) or as hazardous chemical waste.
2. Wipe up spills with paper towel, dispose of with autoclave waste.
3. In event that binding buffer gets in eyes, flush with copious amounts of water at
eyewash station (>15 min), then seek medical attention.
4. If binding buffer contacts skin, wash hands thoroughly and immediately.
5. Any injuries, incidents or near misses (dangerous situations not resulting in an
incident) must be reported to your supervisor and via the online reporting system.
Issue date: 20/11/12
Review date: 20/11/13
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Section 6 – References and Material safety data sheets
1. Read the kit instructions carefully before use, including MSDS for any hazardous
components (typically this is the binding buffer, which is either guanidine
thiocyanate, guanidine HCL, or sodium perchlorate).
2. Risk assessment and SOP “Disposal of chemical waste”
SOP Training Confirmation
By signing below, these individuals confirm that they have read and understood
the SOP, and agree to always follow the instructions in this SOP when performing
this procedure.
Position
Name
Signature
Date
Supervisor
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
employee / student
WHS Committee Approval
Representative: A. Prof Frank Seebacher Chair Safety Committee
Signature: ........................................................ Date: .....................................
Issue date: 20/11/12
Review date: 20/11/13
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