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Prior risk assessment for a technique involving the use of

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									                                                                      MRC Clinical Sciences Centre
                                                                Imperial College Faculty of Medicine
                                                                             Hammersmith Campus
                                                                                     Du Cane Road
                                                                                  London W12 0NN




           RISK ASSESSMENT FOR A LABORATORY PROCEDURE

                                           Technique Title
       Preparing Glycerol stocks of Plasmid DNA


                                            Staff involved
       All Staff in room…………………………


                                          Hazards involved
       Chemicals: Glycerol, LB, Virkon, Ampicillin, Kanamycin.
       Possible exposure to untreated Genetically modified waste
       Possible burns/scalding from Autoclaved waste.
       RSI from pipetting.
       Possible cryogenic burns


                                         Route of exposure
       Skin, eyes, inhalation, ingestion, contact.


                                                Harm
       Chemicals: The respective COSHH forms for the above substances must be read prior to
        this technique commencing so that their hazards and control measures are understood.
       Exposure to GM waste: GM organisms could pose a threat to health.
       Hot waste: Operators could be burned or scalded from the hot contents of the waste
        boxes and liquids on removal from the autoclave. There is also a small risk from steam
       RSI: Long term injury can result from using pipettes and other repetitive techniques.
       Cryogenic burns: Samples stored at -80℃ can burn unprotected skin


 Identify any persons and any special risks associated with the technique and steps that
                           must be taken to minimise them.
       Person carrying out the work: All chemicals, reagents and associated techniques used in
        this protocol will have their associated risks and control measures so their relevant COSHH
        forms should be read in conjunction with this protocol. There is a small risk of exposure to
        viable GMO’s but staff have been trained to work safely with GMO’s and how to properly
        clean up spills should they occur. GMO waste boxes will not be overfilled and so will not be
        excessively heavy. Waste will be allowed to cool after autoclaving to eliminate the risk of
        burns and scalds. RSI can develop from repeated use of pipettes. This needs to be
        monitored and considered for all work. Exposed skin can be burned by racking, samples
        etc. in -80℃ freezers
       Other Persons: Small risk of exposure to chemicals from equipment door handles etc.
        They are only at risk if there is a spill of GMO’s which is very unlikely as these will be
        cleaned up immediately.

       Controls 1 All staff will be given instruction of the hazards and control measures before
        the work commences by an experienced senior member of staff. They will also receive
        appropriate Biological safety and Manual Handling training. Any spillages that occur will be
        cleaned up immediately.
       Controls 2: Appropriate PPE is always worn as well (Nitrile or Latex Gloves and Lab coat,
        Cryogenic gloves). All laboratories have fully functional fume cupboards that are used when
        working with toxic or volatile chemicals. No eating and drinking is allowed in CSC labs

Risk Assessment Form for Laboratory Procedure                                          1
MRC Clinical Sciences Centre
Version 3 July 09
                                                                        MRC Clinical Sciences Centre
                                                                  Imperial College Faculty of Medicine
                                                                               Hammersmith Campus
                                                                                       Du Cane Road
                                                                                    London W12 0NN




 Identify any persons and any special risks associated with the technique and steps that
                           must be taken to minimise them.
        either so there is no real risk of ingestion of chemicals. After the waste has been
        autoclaved it will be allowed to cool before being emptied. The bins will have cooled to
        allow cleaning of the interiors also.


                                  Emergency Spillage Procedure
       All chemicals in the protocol will have the relevant spillage listed in its own COSHH form
        and all staff should familiarise themselves with these procedures.

    For a spill of a GMO:
       Make up 500 ml of 3% Virkon in a beaker, and a sprayer full of 3% Virkon.
       Obtain the Spill kit (should hold posidrive screwdriver, forceps, a clinical waste bag, sharps
        bin and a towel roll).
       Wearing lab coat and gloves spray the 3% Virkon liberally onto the spill.
       Leave it for 30 minutes.
       After 30 minutes; if there is broken glass, use forceps to pick up pieces and place them into
        the empty Sharpsafe. Mop up the spill with the paper towels and put them into a clinical
        waste bag
       Dispose of the clinical waste bag as usual; place any remaining sharps into the sharps bin.
       Inform your supervisor who should either complete a dangerous occurrence form in
        conjunction with the Dept Safety Co-ordinator (or the Divisional Safety Advisor if the DSC
        is away).
       All accidents must be reported to the Lab Manager / Safety Supervisor or the
        departmental first aid personnel, and CSC Safety via an accident report form:
    http://safety.csc.mrc.ac.uk/CSCAccidentReportForm.pdf


                                           Waste Disposal
       Details of the proper means of disposal of chemical waste can be found in individual
        COSHH forms.
       After autoclaving, check the autoclave has cycled properly. If it has dispose of the waste in
        yellow clinical waste bags. Rinse out the autoclave box with tap water and dispose of the
        fluids down the provided sinks.
       Tips eppendorf tubes etc. should all be disposed of as clinical waste.


                     Include a brief stepwise description of the technique
       Put on your laboratory coat and gloves and try to keep within the GM area

               All solid waste must be soaked in a final concentration of 1% Virkon overnight and
                disposed of in the grey autoclave box. Autoclave all waste by the end of the week.
                See (RA for waste disposal”)
               All liquid waste must be made up to at least a final concentration of 1% Virkon
                overnight. Discard down the sink with excess water the following day.
               All glassware and plastics must be soaked in at least a final concentration of 1%
                Virkon for at least 1hour. Rinse thoroughly and place into the washing-up bowls or
                yellow bins as appropriate.

Day One:

       Collect the agar plate containing single colonies of interest
       Inoculate at least 1ml of LB with the appropriate antibiotic with the colony of interest
       Shake at ~200rpm at 37C overnight

Risk Assessment Form for Laboratory Procedure                                            2
MRC Clinical Sciences Centre
Version 3 July 09
                                                                          MRC Clinical Sciences Centre
                                                                    Imperial College Faculty of Medicine
                                                                                 Hammersmith Campus
                                                                                         Du Cane Road
                                                                                      London W12 0NN




                        Include a brief stepwise description of the technique
Day Two:

         Aliquot 150l of sterile glycerol into a cryostat vial
         Aliquot 850l of the fresh overnight culture into the glycerol
         Mix well, label and store at -80C until required


 Due consideration should be made in this assessment for women of child-bearing years, and a full
               re-assessment should be carried out upon notification of pregnancy.

    This assessment has been carried out by the person responsible for the work. A copy of this
                            assessment has been sent to CSC Safety.




RESPONSIBLE PERSON                          NAME                       SIGNATURE               DATE


Person carrying out the
     assessment

 Principal Investigator

 Group Safety Supervisor


        Annual Review




Risk Assessment Form for Laboratory Procedure                                              3
MRC Clinical Sciences Centre
Version 3 July 09

								
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