2.

2 Instrumentation For normal basic functioning of any laboratory, there is need to have effective and necessary equipments (Cheesbrough, 1998)) and the range of instruments available as per each laboratory depends on the different tests carried out. For Research laboratory functioning, some of the required instruments and equipments that were available are listed below: PH meter Vortex mixer Centrifuge Sonicator Ice maker Gel viewing lamp Electric heaters PCR machine Weighing balance Power packs Spectrophotometer ELIZA reader Water distiller Liquid Nitrogen tank Vacuum sealer Shaker Incubator

2.2.1Sonicator This is a machine that utilises both sonic and vibration energy to break the integrity of the cell wall. It was used in the extraction of proteins, it sheers the cell wall thus releasing the cell contents. Principle A Sonicator has a probe connected to a motor that uses electrical energy that causes vibration of the probe at high frequencies; these vibrations cause mechanical shearing of the cell membrane thus releasing the cell contents. The cells are put in a plastic tube with lysis buffer that enhances the disruption. The tube was put on ice so as to reduce heat caused by vibrations that could denature the proteins. Care and Maintenance The machine was placed on a firm bench, prior to use the technologies or students read the SOPs produced by the manufacturers and had to adhere to it. Only skilled personnel handled any technical fault. The Sonicator was always switched off every after use to prevent it from unnecessary exposures to electricity that causes short circuits.

2.2.2 Centrifuge This is an instrument that uses centrifugal force to separate solutions or fluids with different densities. Principle When solutions spanned in a narrow tube, they sediment under relative centrifugal force; depending on their weight, the force applied causes the heavier particles to settle at the bottom of

the tube and less heavier at the top hence forming layers from bottom to top. Centrifugal force applied depends on speed of rotation of the centrifuge rotor induced by the instrument motor. Care and Maintenance Good care was taken to ensure that the machine worked efficiently, prior to use the technologists read the S.O.P provided by the manufacturer and adhered to it, only the manufacturing company serviced the machine. The centrifuge was disinfected and cleaned with 70% ethanol and the exterior cleaned using soap and water for purposes of hygiene. It was essential to balance the number of sample tubes before use, the buckets were checked for any broken particles to avoid excess weight and contamination of the samples. It was only opened when the speed was zero to avoid spills and accidents. For safety purposes only capped tubes were used and not filled to brim to avoid creation of aerosols that could harm the personnel

2.3.2 Spectrophotometer This was a machine used to measure absorbance of solutions basing on Beer Lamberts law, it was used mostly for quantification proteins and nucleic acids. Principle Quantification is based on the amount of light absorbed by the solution containing the substance being quantified. When a beam of light is passed through the solution, some light is absorbed and the other transmitted, the amount of light absorbed is proportional to the concentration of the absorbing substance. The final results are measured in terms of optical density units. Care and maintenance

A spectrophotometer was placed on a firm bench; the electric main was always switched off when not in use and covered with a plastic cover to avoid accumulation of dust that can decrease its sensitivity. For quality results the machines was calibrated before carrying out the quantification by using a blank current. All curvets were washed after use and kept in a dust free cupboard for re-use.

2.2.4 Refrigerator/ Freezers This was an instrument used to provide cold temperature conditions for efficient preservations of samples, reagents and biochemicals in the molecular biology laboratory. Samples, controls, calibrators, and other reagents were stored at low temperatures between 4 to 8C in the fridge and 20- -40C in the freezer section. Principle This equipment operates under the principle that a liquid substance called refrigerant is allowed to circulate in a one-way system between two sets of coiled tubes; One set is the evaporator inside the fridge cabinet and the other is condenser outside the refrigerator, the refrigerant circulates from the compressor to the condenser coil system then to the vaporizer where it is vaporized. During vaporization, latent heat of vaporization is needed and is drawn from the surrounding, the contents of the fridge, and leading to the cooling and freezing effect. The freezers have a high number of coils thus more heat is withdrawn leading to very low temperatures. Care and maintenance The fridge cabinet had a thermometer to monitor the temperature differences and the fridge door was tightly closed whenever not in use to avoid escape of the cool currents. The interior of the

fridge was regularly clean with a light disinfectant like jik to prevent growth of microorganisms; heavy materials were kept at the bottom racks for extra support

2.3 Laboratory layout and safety Safety involves precautions taken in order to protect the work from contamination, equipment from damage, worker and the community from laboratory-acquired infections. Laboratory safety measures that were put in place include laboratory layout and design, personal safety precaution, safe laboratory practices, safe waste disposal, chemical handling and safety precautions against fire. The laboratory was divided into various sections where different activities were done like a separate room for radioactive techniques which was installed with lead shields that prevent the radioactive material from escaping out of the room also had a dosimeter that detects the amount of radioactive material in the room. Designation of specific areas for different procedures reduced contamination and unnecessary spoilages inside the laboratory. The laboratory had two entries that were lockable to prevent unauthorized entry and various glass windows that aided in ventilation and also provided natural light, which was supplemented with electric fluorescent tubes. Fluorescent tubes were preferred because they give better lighting and generate less heat. The laboratory had good supply of running water and electricity the electric wires were sheathed and located on the walls away from the working area and water to avoid accidental running over and electric shocks. The sockets were above the working benches for easy accessibility to the personnel.

Safety was a responsibility of all personnel as the laboratory had many toxic and harmful reagents like ethidium bromide which is highly carcinogenic, thus emphasis was put on personnel protective equipment like laboratory coats, gloves, closed shoes which were worn when carrying out laboratory procedures. Eating, smoking, and applying cosmetics were prohibited in the laboratory sections where infectious materials and tests were carried out except in the conference room. Personnel were encouraged to abide to protocols written down in SOPs on addition guidelines were hanged on the wall to guide the personnel and visitors on how to observe and promote safety in the laboratory. Foods and beverages were not permitted in the laboratory fridges. A sink for hand washing was available and personnel were encouraged to wash their hands on removing gloves and before leaving the laboratory. Politics, sports, conversations were not allowed in the laboratory to boost concentration of the personnel so as to avoid careless mistakes that could become a hazard.

Safe laboratory practices were done to avoid contamination of the personnel with toxic laboratory samples or reagents like mouth pipetting was prohibited to avoid inhalation of reagents that can be toxic. All working benches were cleaned with 10% bleach before and after each working day and in case of a spill it was immediately decontaminated, the benches were made of Formica that is easy to clean and decontaminate. Personnel were discouraged from touching non-infectious materials like door handles, computer with gloves to avoid spread of contamination likewise laboratory coats were prohibited in the conference room.

There was provision of waste buckets for proper disposing of waste materials before incineration and were clearly labelled. Some wastes were decontaminated before disposal like ethidium bromide containing gels. Sweeping was not allowed to avoid dust entering machines so cleaning was done by mopping and whopping of the surface. Contamination of machines with dust would reduce their efficiency due to their high sensitivities. Glassware like slides, test tubes and beakers were immediately washed after use to avoid piling up, they were stored into dust free cabinets.

Reagents and chemicals were handled with care, they were purchased with a material safety data sheet which defined the reagent, safety precautions, ways of handling in case of a spill and disposal procedure and all chemicals were stored according to recommended conditions in the MSDS e.g. flammables away from open flames, keep fuming chemicals covered. Chemicals were labelled clearly with date when prepared or opened. Initials of the technologists and name of the reagents, further more warning labels were put on the containers The laboratory had a bio safety cabinet that was used in handling infectious materials in order to prevent contamination of the work and the worker. A carbondioxide fire extinguisher was also available in case of a fire outbreak.