NSQI LOCAL RULES FOR GENETIC MANIPULATION WORK
CONTENTS:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
Principle Investigators.
Containment Status of NSQI laboratories.
Essential Preliminaries.
Storage of Genetically Modified Material.
Live Cell Working.
Waste Disposal And Treatment Of Contaminated Items For Re-Use.
Spillage.
Centrifugation.
Accidents.
Regulation of Access.
Lab Coats.
1) PRINCIPAL INVESTIGATORS INVOLVED IN BIOLOGICAL WORK AT
NSQI
Monica Berry, Margaret Saunders, Helen Kennedy
2) ACGM CONTAINMENT STATUS AND ORGANISATION OF THE NSQI:
Containment Level
Lab
Group(s)
ACGM
Containment
Level
Wet Lab
Microscope Room
Bacterial Cell
Tissue Culture Lab
Prep Lab 1
Prep Lab 2
Prep Lab 3
2.04
2.04B
2.04A
2.04C
LG.15
LG.14
LG.13
GEN
GEN
GEN
MS, MB
GEN
GEN
HK
1
1
1
2
1
1
2
(GEN indicates communal use.)
LEVEL 2 CONTAINMENT AREAS:
The Wet Lab Tissue Culture (2.04C) and Prep Lab 3 (LG.13) are the only CL-2
areas in the NSQI. Any work requiring ACGM CL-2 containment may ONLY take
place in these rooms. Access is restricted to authorised users only.
All workers intending to use this area must have read and familiarised themselves
with the requirements laid down for CL-2 work in this document and have received
adequate training before hand from suitably qualified and experienced staff.
3) ESSENTIAL PRELIMINARIES
RESEARCH PROJECTS:
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All research projects involving Genetically Modified Organisms (GMO’s) and/or
Containment Level 2 work must be registered with the Safety Office through the host
department. Details of the areas being used to carry out the work in the NSQI MUST
be included in any Risk Assessments associated with that work.
Permission must be obtained by the Principal Investigator for each NEW or
CHANGED project.
NEW WORKERS
Before starting work new workers must:
(a) REGISTER for the work with the University Safety Office (Biological Safety
Officer Dr. Simon Golding, x 88783).
(b) Complete a BIOLOGICAL RISK ASSESSMENT that must be submitted to the
Biological Safety Officer.
(c) RECEIVE TRAINING in handling the GMO’s with which they will be working from
staff fully competent and experienced in genetic manipulation techniques.
(d) READ AND UNDERSTAND:
1) The Biological & Genetic Manipulation Code of Practice (UoB.)
2) The Risk Assessment relevant to the techniques to be used.
3) This document.
4) STORAGE OF GENETICALLY MODIFIED MATERIAL.
STORAGE LOCATIONS:
Fridges, Freezers must be lockable (if sited in a corridor must be locked) and clearly
marked with;
1. a Biohazard Label
2. A label showing the Classification of the material stored e.g.GMM1.
3. Contents list or Reference to the location where contents list is held
INVENTORY: The workers concerned in each group are responsible for ensuring
that lists are updated each time a new GMO is added to the inventory.
5) LIVE CELL WORKING
LOCATION:
CL-1:
Bacterial Cell Culture (2.04A)
Microscope Room (2.04B)
Main Wet lab area (2.04)
Basement Prep Labs (LG.14 & 15) & All Low Noise Labs
CL-2: Restricted to Wet Lab Tissue Culture (2.04C) & Prep Lab 3 (LG.13)
RULES:
2
(1.)
Labcoat, safety goggles disposable gloves must be worn when working in
these areas.
(2.)
Surfaces must be swabbed with ethanol (70%) after EACH use and left clear.
(3.)
The working area must be maintained in a CLEAN and TIDY state. No
materials should be left cluttering the surface after use.
(4.)
Wash bottles containing Ethanol (70%) or Chloros (Sodium Hypochlorite;
10%) must ALWAYS be present on the bench. Stocks of Virkon should be
available. Chlorhexidine wipes may also be used for surface disinfection.
(Neat Chloros should be available for dealing with large volume spills).
(5.)
The Buchner flask attached to the aspiration pump in the Tissue Culture Lab
must always contain FRESH CHLOROS.
(6.)
Major spillages must be reported immediately to Stuart Bellamy and Fred
Hale, the SSA. Emergency advice can be obtained from the Safety Office.
(7.)
Staff must ensure that they know the location of the nearest spillage capture
kit and granules and approved contingency plans for dealing with spillage
BEFORE undertaking any work (see Spillage).
(8.)
Instructions displayed in CL-2 Tissue Cultures must be followed.
(9.)
Centrifuge tubes/bottles should only be filled on Spill Trays.
N.B. Sodium Hypochlorite (trade name Chloros) is provided as 10-11% solution, so
no need to dilute. All additions of Chloros to cell waste/washings should be to a final
concentration of at least 1% v/v (1:10 dilution).
6) WASTE DISPOSAL AND TREATMENT OF CONTAMINATED ITEMS
FOR REUSE
SOLID BIOHAZRAD WASTE
This should be placed in a yellow lab bin (these are suitable for Biohazard waste).
When no more than 2/3 full the bag should be securely tied (double bag if
necessary) and placed next to the front entrance of the wet lab.
LIQUID BIOHAZARD WASTE
Liquid waste is dealt with according to the vessel in which it is contained:
(1.)
In 15 or 50 ml falcon tubes: Ensure lid is securely fastened and dispose of
in a yellow lab bin
(2.)
In any glassware: add Chloros, leave overnight and dispose of liquid down
the sink with copious amounts of water. Do not dispose of during heavy rain
to prevent drain overspill. Record the amount of liquid disposed of (cell
volume and total volume). Glassware can then be washed for re-use.
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PLEASE NOTE THAT WASTE CONTAINING NANOPARTICLES MUST BE
DIPOSED OF BY A SEPARATE ROUTE.
WASTE DISPOSAL: SPECIAL PROVISION CL-2 AREAS
The disposal of waste generated in a CL-2 area must be assessed on a case by
case scenario, guided by risk assessment. Most CL-2 waste can be contained and
placed into the yellow lab bins, however some CL-2 waste must be autoclaved. The
NSQI has an in-house autoclave located in the LG.13 for this purpose. In order to
minimize the volume of such waste, all consumables to be used in a CL-2 room
should be unpacked before taking them into the room so that the packaging can be
disposed of by the normal route for non-hazardous material.
DECONTAMINATION OF RE-USABLE ITEMS NOT SUITABLE FOR
AUTOCLAVING: NB. All re-usable items other than those listed below must be
autoclaved before washing and re-use.
Glass Pipettes
These can be decontaminated by complete immersion in 1% chloros overnight.
Centrifuge Tubes
Preparation for re-use:

Supernatants (e.g. contaminated growth media) must be decontaminated with
Chloros before disposal down the sink with copious amounts of water.

If pellets are being discarded they should be resuspended and poured into the
same vessel with subsequent rinsings and treated as above.

Tubes and bottles containing waste should then be totally immersed in 1%
chloros for a minimum of 24 hrs. If the tubes or bottles are incompatible with
chloros, follow manufacturer’s instructions for decontamination.

Before transferring to the normal washing up, chloros from the above should be
disposed of via the laboratory sink and tubes & bottles thoroughly rinsed with
copious amounts of tap water.
Buchner Flasks Used Exclusively to Collect Microbiologically Contaminated,
Aspirated fluids.
Initially, sufficient 10% Chloros should be added such that the final concentration of
Chloros is between 1-5% v/v. When all of the aspirant has been collected, ensure
that it is thoroughly mixed with the Chloros. Let it stand for 24hrs, then pour the
liquid down a lab sink. The flask can then be rinsed out and re-used for collection of
supernatant. Record the amount of liquid disposed of (cell volume and total volume).
ETHIDIUM BROMIDE WASTE
1. Soft Waste: inc. gels
4
Place ethidium stained gels in a small white bucket located under the gel running
area (this is clearly labelled for gels). When ¾ full place lid on bucket and label with:
Lab, Date, ‘Ethidium Bromide Gel Waste’, ‘CARCINOGENIC’. These will then be
disposed of by Stuart Bellamy (sent for incineration).
2. Plastic & Tissue Waste: inc. tips, tubes, vials etc.
Put into ‘Disposafe’ plastic screw top plastic jar (‘sweety jar’). Fill until ¾ full, seal
with parafilm and label with Lab, Date, ‘Ethidium Bromide Plastic & Tissue Waste’,
‘CARCINOGENIC’. These will be then be disposed of by Stuart Bellamy (sent for
incineration).
3. Sharps: inc. needles, blades, glass pasteurs.
Rinse thoroughly with water and dispose of through normal lab waste (‘Sharps
Waste’ for needles & blades; ‘Glass Waste’ for glass pasteurs [both sealable yellow
cinbins]).
4. Syringes
Again rinse thoroughly with water and dispose in normal ‘Syringe Waste’ (sealable
yellow cinbins).
7) SPILLAGE
ACTION IN RESPONSE TO SPILLS: All workers must be aware of the correct
course of action in response to spillage of microbiological material. This is
summarised in the Table below.
Type of Spill
On bench
Action
a) Absorb spill with paper tissues and then wipe bench
surface with tissues impregnated with chloros (10%) or
ethanol (70%)
b) OR
wipe
the
area
with
a
Wipex
Cloth
equivalent.(Chlorhexidine Wipe)
c) Dispose of all waste in yellow lab bins.
On Floor (small-scale)
As above
On Floor (large-scale)
a) Warn others to keep away from area affected
b) Absorb the spill using Trivorex spillage granules and
surround the spill with absorbant pads
c) Dispose of pads in a yellow lab bin.
d) Wipe area with tissues soaked in chloros (10%) or
ethanol (70%)
e) Dispose of tissues in a yellow lab bin.
On Labcoat
Remove lab coat immediately and place in a Biohazard bag
for subsequent Autoclaving before putting into normal
laundry system (See ‘Laundry Arrangements’)
On Personal Clothing or
Treat as for Labcoats. One size Disposable Coverall Suits
Footwear
(kept on shelf at entrance of Wet Lab) are available for use
in emergency use if personal clothing becomes
contaminated. Do not leave the lab wearing contaminated
clothing
On exposed Skin or Hair Wash affected area repeatedly with soap and water.
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In Centrifuge bowl or
rotor
In incubator
On Broken Glass (smallscale)
On Broken Glass (largescale)
Consult the SSA/BSO for advice.
Mop up spill with tissues and swab with Ethanol (70%).
Place all waste in a yellow lab bin
Turn off incubator
Check for broken glass
Mop up spillage with paper tissues.
Swab all affected surface with chloros (10%) or ethanol
(70%).
N.B. It may be necessary to remove the bottom support
tray. When the spillage has been removed, swab surfaces
with Chloros/Ethanol. Dispose of tissue waste in a yellow
lab bin
Treat with great caution. Place glass fragments in a
container and immerse in 1% chloros for at 24 hours to
decontaminate.
Alert Stuart Bellamy and/or SSA immediately to discuss
decontamination and disposal.
N.B. Serious large-scale spillage incidents e.g. where contaminating fluids run under
benches, must be recorded in the lab Incident Report Book (located on the shelf with
the Safety Documents in Technicians Office). The SSA (Fred Hale) and/or the
University Biological Safety Officer (BSO: Dr.S.Golding X 88783) should be
consulted for advice and to check that the all necessary actions have been taken to
render the area safe for reuse.
SPILLAGE: SPECIAL PROVISION CL-2 AREAS
If an unidentified spill is discovered in a CL-2 area, do NOT attempt to clean-up and
contact Stuart Bellamy immediately. The spill may contain agents harmful to human
health, such as Hepatitis B.
8) CENTRIFUGATION
GOOD PRACTICE
(1.)
Carefully check tubes and bottles for cracks, discoloration and other damage
prior to use. Do not use if in doubt.
(2.)
Fill tubes/bottles according to manufacturer’s instructions on a spill tray and
balance carefully.
(3.)
Swab centrifuge bowls and rotor pockets with Ethanol (70%) after use.
(4.)
Decontaminate tubes and bottles before washing according manufacturers
guidelines.
TRANSPORTING CENTRIFUGE TUBES AND BOTTLES:
Secondary containment must be employed when transporting centrifuge tubes and
bottles outside the laboratory. Bottles should be sealed inside a sturdy sealed
polythene container and carried in a bucket or specialised rack to prevent tipping.
9) ACCIDENTS
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NOTIFICATION
Serious accidents must be reported immediately to the University Safety Office
(x88780) and to the SSA (Fred Hale x40004), who will decide what action is
required.
Minor accidents may be dealt with at the departmental level initially after consulting
the SSA.
ACCIDENT REPORT FORM
All
accidents
must
be
recorded
on
an
Accident
Report
Form
(https://www.bris.ac.uk/safety/accidents.html). The form must be passed on to the
NSQI SSA and host Head of School as soon as possible so that it can be forwarded
to the Safety Office within 3 days of the occurrence.
FIRST AID
If personal injury is involved First Aid may be rendered by University Approved First
Aiders. For the NSQI, these are Fred Hale, the SSA, Dr Monica Berry and Stuart
Bellamy, the wet lab manager. Cuts and scratches which have come into contact
with microorganisms should be swabbed with anti-bacterial wipes (if available).
HOSPITAL ATTENDANCE
This is not mandatory. If hospital attendance is necessary and the injury has involved
exposure of a wound to GMO’s, the hospital should be told of any antibiotic
resistance possessed by the bacterial strain involved. However, it is not a
requirement to mention that recombinant material was involved.
FOLLOW UP
Wounds which have been contaminated as above must be closely monitored for
signs of infection and medical advice sought immediately if any signs become
apparent.
10) REGULATION OF ACCESS
RESTRICTION OF ENTRY
Access to the lab must be strictly regulated for all non-lab staff. Visitors with
legitimate business in the lab should be carefully supervised and confined to areas
outside the restricted areas of the lab unless entry to the restricted area is essential
to carry out their work. In order to gain entry they MUST sign in on the form at
reception and obtain a visitor’s pass, which can be issued by a one of the NSQI lab
managers.
NON-LAB STAFF
This includes personnel such as Service Engineers, Salesmen and Technical reps
who come to demonstrate apparatus and Bursars Office Staff e.g. Plumbers,
Electricians etc.
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CLEANERS
Room cleaning is normally carried out by lab staff. Housekeeping staff carry out floor
cleaning; in order for them to carry this out, the floor area must be cleared of
obstructions (e.g. chairs) and decomtaminated.
CL-2 LEVEL LAB ACCESS
This should be restricted to authorised users only, identified by way of a completed
and signed risk assessment. UNDER NO CIRCUMSTANCES should visitors or
unauthorised lab users enter these areas. Cleaning staff are also NOT permitted to
enter any CL-2 area.
11) LAB COATS
CL-1 CONTAINMENT AREAS: White Lab coats
RULES:
Lab coats must be worn at all times when carrying out practical work
within the lab. Lab coats must not be worn in the Writing Up areas or
offices.
Coatpegs
Lab coats must be hung on coat pegs marked “Labcoats only” when
not in use and MUST be kept separate from outside coats. They must
never be left on chairs.
Laundry
Soiled Labcoats should be placed in the Blue Bag under the bench of
Stuart Bellamy.
CL-2 CONTAINMENT AREAS: Blue Lab coats
These coats must be worn when working in the CL-2 area and removed and left
inside the CL-2 Lab on leaving. Coats must be autoclaved before laundering. Special
laundry arrangements are adopted for these coats.
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