Laboratory Safety Procedures for Molecular biology lab.

A. GENERAL B. SHARP OBJECTS AND BROKEN GLASS E. INSTRUMENTS AND EQUIPMENT G. BODY FLUIDS

A. GENERAL 1. Work carefully and cautiously in the laboratory, using common sense and good judgment at all times. 2. EATING AND DRINKING ARE PROHIBITED in the laboratory. 3. Long hair must be tied back during laboratory sessions. 4. Identify the location of all exits from the laboratory and from the building. 5. Keep sinks free of paper or any debris that could interfere with drainage. 6. Lab tables must be clear of all items that are not necessary for the lab exercise. 7. Wash hands and the lab tables with the appropriate cleaning agents before and after every laboratory session.

B. SHARP OBJECTS AND BROKEN GLASS 1. Pointed dissection probes, scalpels, razor blades, scissors, and microtome knives must be used with great care, and placed in a safe position when not in use. 2. Containers designated for the disposal of sharps (scalpel blades, razor blades, needles; dissection pins, etc.) and containers designated for broken glass are present in each laboratory. Never dispose of any sharp object in the regular trash containers. 3. Do not touch broken glass with bare hands. Put on gloves and use a broom and dustpan to clean up glass. Dispose of ALL broken glass in the specific container marked for glass. Do not place broken glass in the regular trash.

E. INSTRUMENTS AND EQUIPMENT Care must be used when handling any equipment in the laboratory. Students are responsible for being familiar with and following correct safety practices for all instruments and equipment used in the laboratory.

G. BODY FLUIDS Special precautions are to be followed in all laboratories using any body fluids, such as blood, saliva, and urine, because of the potential to transmit disease-causing organisms. 1. Use gloves in all laboratory experiments that involve the use of body fluids. 2. All contaminated material, such as slides, coverslips, toothpicks, lancets, alcohol swabs, etc., must be placed in a biohazard bag for proper disposal and should never be reused.

LAB EQUIPMENTS

1-Refrigerated Centrifuge : Function: Separates different components of a volume of samples by means of a centrifugal force based on their differing density. Purpose: Enable the collection of desired components that had been separated. The adjustable temperatures allow operator to choose the desired temperature to work with their samples (this is especially useful for heat sensitive sample)

2-Micropipette : Function: Micropipette are the standard laboratory equipment used to measure and transfer small volumes of liquids Purpose: Each pipet can be set to transfer any volume within its own volume range.

Three sizes of micropipettes The micropipettors in this laboratory come in three different sizes each of which measures a different range of volumes. The three sizes are P20, P200 and P1000. These sizes are noted on the top of the plunger button. Size Micropipette Range of volumes measured P μl P μl P μl

Eppendorf tubes DISPOSABLE TIP. Disposable Transfer Pipettes disposable serological pipettes

3-Lab Companion Shaker : Function: its shaking ability to create aeration and homogeneity in the sample. Purpose: provide a homogenise samples and the adjustable temperatures allow operator to choose the desired temperature to work with their samples (this is especially useful for heat sensitive sample) 4-Sensitive Balance: Function: To weigh out small samples with readings up to three decimal place.

5-Water Bath : Function: provide a steady temperature for incubation of solution. Purpose: Ensure that such solutions equilibrate to a particular temperature for and during use. 6-Ice maker Machine: Function: Production of ice flakes to be stored in the storage chamber. Purpose: the ice is used to keep samples or enzymes cool to prevent them from becoming denatured.

7-Autoclave : Function: Sterilise equipment and solution by killing micro-organisms with high temperature and steams. Purpose: Prevents contamination Autoclave tape: A roll of 19mm x 55m tape used to secure packages going into an autoclave (with a vacuum cycle) which changes colour to indicate sterilisation procedure is complete.

8-Laboratory Refrigerator: function: manufactured to cool down the temperature. Purpose: The adjustable temperatures allow operator to choose the desired temperature to keep their samples fresh.

Blood Collection Tubes

EDTA TUBES: EDTA tube is widely used for clinical hematology as well as various kinds of blood cell test instrument. It makes use of EDTA as anticoagulant. Meanwhile, it offers a comprehensive protection for blood cell, especially for protecting the blood platelet, so that it can effectively stop the gathering of blood platelet and makes the form and volume of blood cell uninfluenced in a long time. EDTA tube can be used for DNA extraction and plasma isolation.

HEPARIN TUBES: Heparin tube is used for blood collection and anti- coagulation, not only for routine clinical biochemistry tests and emergency biochemistry tests, but also for some test items in blood theology. It is coated with heparin. The anticoagulant heparin activates anti-thrombins, thus blocks the coagulation cascade and produces a whole blood/ plasma sample instead of clotted blood plus serum. In addition, this type of tubes can be used for cell culture

NO ADDITIVE TUBES: No additive tube is used for blood collection and storage for biochemistry, immunology and serology tests in medical inspection. It can provide enough and non-polluted serum specimen for clinical tests, while keeping the serum invariable in the long inspection period. It is applicable for all current mainstream biochemical analyzers

Isolation of Serum and Plasma

Note: Blood Plasma contains many vital proteins including fibrinogen, globulins and human serum albumin. Serum refers to blood plasma in which clotting factors (such as fibrin) have been removed naturally by allowing the blood to clot prior to isolating the liquid component - Isolation of the Plasma from Blood. 1. Collect the blood in EDTA (purple top) tubes. 2. Centrifuge at 3000 rpm for 15 minutes. 3. Transfer the Plasma to 1.5 ml tubes(-20).