LBL02001: Difference between revisions

'''6.1 Laboratory Access Control'''

The lab can be accessed using the number lock or using the card sensor to unlock the door, if your access card (such as an oyster card) has been registered.

Access control to the laboratory area is to ensure that only trained and responsible individuals can make use of the laboratory facilities of which this risk assessment is primarily concerned.  

'''6.2 PPE, Biosafety and Contamination Risks'''

The kind of biological organisms with which we typically work in the laboratory include

i) non-pathogenic strains of E. coli (such as DH5 alpha),

'''6.3 Microwave'''

The microwave is a familiar device to probably all laboratory users and the standard precautions apply when using it. In particular, the handling of hot liquids and glassware may present the risk of burns and as such thermally resistant gloves are provided. Of note is also the fact that one of the primary uses of the microwave is heating of agarose for the casting of electrophoresis gels. User’s should be aware that previous, possibly less careful, operators may have used the microwave in conjunction with biocontaminants and/or hazardous chemicals (as described later) and as such operation of the microwave using gloves and goggles is recommended in any case.

[associated SOPs]

'''6.4 Spectrophotometer'''

The spectrophotometer is a device that measure the absorbance of a given liquid sample at various wavelengths of light. A full description of it’s usage is given in the SOPs [SOP]. User’s should be aware that this system can generate light of significant intensity in both ultraviolet and infrared wavelengths. While most of the optical path and the bulbs required to generate this light is contained within the system, the user may come into contact with this in the sample measurement compartment of the device. As such it should be operated only with the sample measurement comparment fully closed.

'''6.5 PCR Machine'''

The Polymerase Chain Reaction (PCR) machine, also known as a thermalcycler, is used to amplified specific sections of template DNA either for diagnostic or molecular cloning purposes. The heating block and underside lid of this machine will typically reach temperatures in the region of 95 to 100degC in a typical amplification program and caution should be exercised when placing and removing samples. Information regarding the current state of the machine including current program and temperature is displayed on the LCD on the front panel of this machine. Full information regarding the use and programming of the Techne thermal cycler and it's use in various technical can be found in the SOPs and technical documentation. [SOPs, docs]

'''6.6 Electrophoresis Power Supply Unit'''

The Power Supply Unit (PSU) used to supply a current to the electrophoresis tank (described below) is capable of producing dangerously high voltages and currents. It is typically set to the levels required for DNA electrophoresis in 100mL 1 - 2% w/v agarose gels. Caution should be exercised when plugging in and handling electrodes due to risk of electric shock. Use of a residual current device is recommended when using this or any other electrical device in the laboratory that may pose a risk to operators from electric shock. This instrument is also often used in conjunction with Ethidium Bromide based procedures (see below) and should be operated using gloves.

'''6.7 Electrophoresis tank and Ethidium Bromide area'''

The electrophoresis tank is located within the electrophoresis/Ethidium Bromide area of the laboratory bench. It is advised any work carried out within this area is manipulated separately from any work done in other areas. Any consumables used when handling Ethidium Bromide such as gloves and tips should be disposed of before handling anything outside of this area to prevent contamination of the wider lab area with the potentially toxic chemicals used in DNA staining. Separate pipets designated for ethidium bromide work and labelled as such should only be used for molecular biology work in this area and these pipets should not be used anywhere else. A full description of how to set up and run agarose gels for DNA electrophoresis is described in the SOPs [SOP].

'''6.8 UV illuminator'''

Gel electrophoresis separates DNA by size within an electric field. Separation of varying sizes of DNA fragment within an agarose gel allows visualisation of the various sizes of DNA fragment contained within a sample when nucleic acid binding stains are used. The typical stain used is Ethidium Bromide which is a fluorophore that is excited within the UV spectrum (with excitation maxima under 300nm) and emission within the visible spectrum. This presents two main risks to the user:  

i) As a DNA intercalating agent Ethidium Bromide is potentially carcinogenic and should never be allowed to come in contact with the user’s skin, caution should therefore be applied throughout the entire DNA electrophoresis procedure from gel preparation, through electrophoresis and then subsequent gel visualisation. This is therefore also carried out in the electrophoresis/ethidium bromide area of the laboratory bench.

'''6.9 HEPA flow cabinet/area'''

The HEPA filtered laminar flow unit allows us to work in sterile air in order to prevent contamination of our work such as petri dishes and broths with other microorganisms. Proper usage of the laminar flow area is described in the SOPs. It should be noted that the laminar flow functions in such a way as to protect the user’s work rather than the user and so caution still be taken by the user to maintain the aseptic conditions of anything used within the flow area and in disposal of consumables to assist in containment of biomaterial. See SOPs on aseptic technique. [SOP].

'''6.10 Hot plate/Steam cooker/steriliser/autoclave'''

The sterilisation unit [autoclave?] is used to destroy any potential microorganisms that might contaminate media, reagents and consumables to be used aseptically in micro and molecular biological procedures. High temperature and pressure is used to kill contaminants and the main risk to the user is from the heat of the metal pressurised unit during sterilisation and any vented steam. The system itself is unpowered and is heated using the hot plate which itself becomes hot enough to cause serious burns. Thermal gloves are available for handling the autoclave and material processed by it. It is recommended that the autoclave and contents are allowed to cool for a while before handling. The correct procedures for using the autoclave/steriliser to prepare media and destroy GM waste are described in the SOPs. [SOPs]

'''6.11 Sink area'''

The sink area should remain clear. All glassware should be kept clean and out of the way.

'''6.12 Chemicals and Storage'''

As per chemical list, and MSDS by chemical [SOPs appendix A]

'''6.13 Incubator'''

While the incubator itself does not pose a high risk due to typically operating in the temperature required for the culture of mesophiles. care should be taken when moving samples to and from the incubator as per handling biocontaminants mentioned previously. Many procedures described later in the SOP documentation indicate the need for incubation and details can be located there. [SOPs]

'''6.14 Centrifuges'''

The laboratory has a number of centrifuges available for use.  

The Jouan is a larger device which can take 50ml tubes and larger containers if fitted with buckets and can spin up to 10,000 rpm (although documentation and instrument panel indicate higher rpm may be possible this particular unit appears to have a limit of 10,000.