NSQI LNL local rules for genetic manipulation work
Contents
1. Principal investigators involved in biological work at NSQI LNL ..................................................... 2
2. ACGM containment status and organisation of the NSQI LNL ....................................................... 2
Level 2 containment areas .............................................................................................................. 2
3. Essential preliminaries .................................................................................................................... 2
Research projects:........................................................................................................................... 2
New workers ................................................................................................................................... 2
4. Storage of genetically modified material ........................................................................................ 3
Storage locations: ........................................................................................................................... 3
Inventory ......................................................................................................................................... 3
5. Live cell working .............................................................................................................................. 3
Location........................................................................................................................................... 3
Rules ................................................................................................................................................ 3
6. Waste disposal and treatment of contaminated items for reuse................................................... 4
Solid biohazard waste ..................................................................................................................... 4
Liquid biohazard waste ................................................................................................................... 4
Waste disposal: special provision CL-2 areas.................................................................................. 4
Decontamination of re-usable items not suitable for autoclaving ................................................. 4
Ethidium bromide waste ................................................................................................................. 5
7. Spillage ............................................................................................................................................ 5
Action in response to spills: ............................................................................................................ 5
Spillage: special provision CL-2 areas.............................................................................................. 6
8. Centrifugation ................................................................................................................................. 6
Good practice .................................................................................................................................. 6
Transporting centrifuge tubes and bottles: .................................................................................... 6
2. Accidents ......................................................................................................................................... 6
Notification ..................................................................................................................................... 6
Accident report form ...................................................................................................................... 7
First aid............................................................................................................................................ 7
Hospital attendance ........................................................................................................................ 7
Follow up......................................................................................................................................... 7
10. Regulation of access...................................................................................................................... 7
Restriction of entry ......................................................................................................................... 7
Non-lab staff ................................................................................................................................... 7
Cleaners........................................................................................................................................... 7
CL-2 level lab access ........................................................................................................................ 7
11. Lab coats ....................................................................................................................................... 7
CL-1 containment areas: White Lab coats ...................................................................................... 7
CL-2 containment areas: Blue Lab coats ......................................................................................... 8
1. Principal investigators involved in biological work at NSQI LNL
Currently none
2. ACGM containment status and organisation of the NSQI LNL
Lab space
Lab
Wet Lab
2.04
Microscope Room
2.04B
Bacterial Cell
2.04A
Tissue Culture Lab
2.04C
Prep Lab 1
LG.15
Prep Lab 2
LG.14
Prep Lab 3
LG.13
(GEN indicates communal use.)
Group(s)
GEN
GEN
GEN
MS, MB
GEN
GEN
GEN
ACGM Containment
Level
Decommissioned
Decommissioned
Decommissioned
Decommissioned
1
1
1
Level 2 containment areas
The 2.04C and LG.13 were originally designated as CL-2 areas in the NSQI. Since then the 2.04
complex has been decommissioned and LG.13 has been downgraded to CL-1. Any work requiring
ACGM CL-2 containment would require assessment and redesignation of lab space.
All workers intending to use CL-2 space 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:
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 LNL 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:
1. Register for the work with the University Safety Office (Biological Safety Officer Dr. Simon
Golding, x 88783).
2. Complete a Biological risk assessment that must be submitted to the Biological Safety
Officer.
3. Receive training in handling the GMO’s with which they will be working from staff fully
competent and experienced in genetic manipulation techniques.
4. Read and understand:
 The Biological & Genetic Manipulation Code of Practice (UoB.)
 The Risk Assessment relevant to the techniques to be used.
 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.
2.
3.
A Biohazard Label
A label showing the Classification of the material stored, e.g. GMM1.
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:


Basement Prep Labs (LG.13, 14 & 15)
All Low Noise Labs
CL-2:

Currently no designated space
Rules:
1.
2.
3.
4.
5.
6.
Labcoat, safety goggles disposable gloves must be worn when working in these areas.
Surfaces must be swabbed with ethanol (70%) after EACH use and left clear.
The working area must be maintained in a CLEAN and TIDY state. No materials should be
left cluttering the surface after use.
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).
The Buchner flask attached to the aspiration pump for Tissue Culture must always
contain FRESH CHLOROS.
Major spillages must be reported immediately to the Lab manager or the Centre
manager. Emergency advice can be obtained from the Safety Office.
7.
8.
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).
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 biohazard 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.
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.
Decontamination of re-usable items not suitable for autoclaving
1. Glass Pipettes
These can be decontaminated by complete immersion in 1% chloros overnight.
2. 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.
3. 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
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 the Lab
manager (sent for incineration).
1. 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 the Lab manager (sent for
incineration).
2. 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
On Floor (small-scale)
On Floor (large-scale)
On Labcoat
On Personal Clothing or
Footwear
On exposed Skin or Hair
Action
1. Absorb spill with paper tissues and then wipe bench
surface with tissues impregnated with chloros (10%) or
ethanol (70%)
2. OR wipe the area with a Wipex Cloth
equivalent.(Chlorhexidine Wipe)
3. Dispose of all waste in yellow lab bins.
As above
1. Warn others to keep away from area affected
2. Absorb the spill using Trivorex spillage granules and
surround the spill with absorbant pads
3. Dispose of pads in a yellow lab bin.
4. Wipe area with tissues soaked in chloros (10%) or ethanol
(70%)
5. Dispose of tissues in a yellow lab bin.
Remove lab coat immediately and place in a Biohazard bag for
subsequent Autoclaving before putting into normal laundry
system (See ‘Laundry Arrangements’)
Treat as for Labcoats. One size Disposable Coverall Suits (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
Wash affected area repeatedly with soap and water. Consult the
SSA/BSO for advice.
In Centrifuge bowl or rotor
Mop up spill with tissues and swab with Ethanol (70%). Place all
waste in a yellow lab bin
In incubator
1. Turn off incubator
2. Check for broken glass
3. Mop up spillage with paper tissues.
4. Swab all affected surface with chloros (10%) or ethanol
(70%).
5. 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
On Broken Glass (smallTreat with great caution. Place glass fragments in a container and
scale)
immerse in 1% chloros for at 24 hours to decontaminate.
On Broken Glass (large-scale) Alert the lab manager 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, x88783) 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 the Lab
manager 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 manufacturer’s 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
Notification
Serious accidents must be reported immediately to the University Safety Office (x88780) and to the
SSA (Fred Hale x46953), 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 Peter Dunton and Alex Martin. 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 LNL 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.
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
decontaminated.
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.


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.
Soiled Labcoats should be brought to the attention of the Lab manager.
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.