"Prior risk assessment for a technique involving the use of - Get Now DOC"
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 Sequencing PCR or DNA samples Staff involved All Staff in room……………………… Hazards involved Chemicals: Ethidium Bromide, DNA Loading dye: (Bromophenol blue, EDTA, Xylene cyanol). Possible electric shock. UV light burns Sharps injury RSI from pipetting Route of exposure Skin, eyes, 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. Electric shocks: can burn and in some cases kill, especially when used near liquid or if faulty equipment is used. UV light: Can burn skin as well as damage the eyes. Repeated exposure could lead to irreversible effects. Sharps: Razor blades/scalpels can cut very deeply and can also introduce contaminants into a wound RSI: Long term injury can result from using pipettes and other repetitive techniques. 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. UV can burn exposed skin, the face and damage the eyes. Power packs can potentially cause electric shocks. Scalpels and razor blades need to be used very carefully. RSI can develop from repeated use of pipettes. This needs to be monitored and considered for all work. Other Persons: Small risk of exposure to chemicals from equipment door handles etc. Small risk of UV burn if they are too close to the gel imager while it is switched on. Electrical hazard from the gel tanks and power packs. Risk of sharps injury if scalpels etc. have not been disposed of properly. Controls 1: All new staff and students have a laboratory induction to ensure that they understand all of the safety considerations associated with the work carried out within this laboratory. All techniques are explained and demonstrated to all new staff and students. Staff are sent on the appropriate training courses offered by the MRC to ensure they are familiar with all aspects of good laboratory practice. Controls 2: Appropriate PPE is always worn as well (Nitrile or Latex Gloves and Lab coat 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. etc. Face protection needs to be worn when working with UV light.). 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 either so there is no real risk of ingestion of chemicals. All electrical equipment is PA tested on a bi-annual basis and all new equipment is PA tested when it arrives in the department. 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. All other relevant Risk assessments associated with this protocol should also be read. 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. Tips eppendorf tubes etc. should all be disposed of as clinical waste. Ethidium Bromide solid waste needs to be placed into the special “for incineration only” bin Ethidium Bromide liquid waste needs to be cleaned up either using the purification column or “Tea Bag” method. Include a brief stepwise description of the technique Put on your laboratory coat and gloves Collect your prepared agarose gel (See RA for making an agarose gel’ sheet) Load the PCR reaction or DNA sample to be sequenced into one of the lanes of the gel (See ‘RA for loading and visualising DNA bands on an agarose gel’ sheet) Run the samples until enough separation is gained Take a picture of the resulting DNA bands (See ‘RA for loading and visualising DNA bands on an agarose gel’ sheet) Cut out the band to be sequenced, trimming off as much excess agarose as possible (See ‘RA for gel extracting DNA fragments’ sheet) Resuspend the DNA in as small a volume of buffer as possible (usually 30l) Estimate the amount of DNA per sample using the Taq/Pvu markers as a guide (see below) Marker size (bp) DNA concentration 1443 135ng 1008 94ng 613 57ng 357 33ng 278 26ng 193 18ng 108 10ng per 15l of 25ng/l Taq/Pvu marker Collect and label a 1.5ml eppendorf Aliquot 1l of 25ng/l (or 10M stock solution) of appropriate primer into the eppendorf Aliquot approximately 350ng of DNA sample per 3Kb of DNA size Add the appropriate amount of dH2O to make a final volume of 10l Fill out a sequencing request form and get an ordering number 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 Take the samples to the DNA core unit in the hospital In a couple of days, check your DNA folder via fetch on the Mac for the results Analyse the results on the Mac using the editview, DNAstrider and Netscape programs 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