UNIVERSITI MALAYSIA SABAH SCHOOL OF INTERNATIONAL TROPICAL FORESTRY

PATOLOGY PRACTCLE: 1 TITLE: MEDIA PREPARATION LECTURER: ASSOCIATED PROFESSOR DR. MAHMUD SUDIN NAME: YATCHI CHONG MATRIX NO.: BF11110173 DATE: 1/3/2012

Title: Media Preparation Objective: - To give exposure to students in terms preparing commercial media such as MEA (Malt Extract Agar) - Emphasizing on the right technique of media preparation without contamination or defects. - Students are also exposed to variety of laboratory instruments to ease the laboratory work or media preparation. Material: - MEA (Malt Extract Agar) Apparatus: - Spatula, - Weighing boat, - Petri dish, - Parafilm, - Analytical Balance, - 250ml Duran bottle, - Autoclave machine, - Marking tape, - Heat source (candle/lamp), - Sanitizer, - Laminar flow chamber. Procedure: 1. Measure the weight of the weighing boat on the analytical balance and set it to zero. 2. Use spatula to take 12g of MEA into the weighing boat. 3. Then pour the MEA into the 250ml Duran bottle. 4. Fill in the Duran bottle with distilled water till it reached 250ml. 5. Shake the bottle until the MEA fully dissolve. 6. Loosen the bottles cap a little (one or two spin), then put the marking tape on the cape till the glass part of the Duran bottle (enough to hold/stabilized the cap). 7. Put the Duran bottle on the provided fences then it into the autoclave machine. 8. Set the autoclave machine to 121C for 20 minutes. 9. After 20 minutes, wait for the Duran bottle to cool down about 40C. 10. Then take the Duran bottle into the laminar flow chamber. 11. Pour some sanitizer on your hand to prevent any contamination on the apparatus within the laminar flow chamber. 12. Hold both Duran bottle and Petri dish without touch it on the laminar flow chamber base. 13. Open the Petri dish and hold it against the heat source (lamp). To kill of any contaminant on the Petri dish.

14. Then pour the MEA into the Petri dish (fill in about quarter of the Petri dish and distribute it evenly). 15. Leave the Petri dish (opened) within the laminar flow chamber for it to cool down and solidify the MEA. 16. Then seal it with the Parafilm and label it. Result: - 3 media is successfully prepared. Discussion: - Autoclave machine; o Fill in distilled water into the exhaust bottle within the minimum and maximum range (mark on the bottle), fill it about 70% of the bottles volume or according to how many Duran bottle placed within autoclave. o Shake the exhaust bottle to check to check whether there is leakage upon the bottle then closed the lid tightly and connect it to exhaust host. o Then set the autoclave machine to 121C for 20 minutes. o If the temperature is lower the process and time take n for the process might takes even longer than 20 minutes. o If the temperature is too high, there is possibility that the Duran bottles cap might melt. Although the time taken for the process is shorter. o The purpose of the autoclave machine is too steam as well as to exterminate any contamination on the Duran bottle. Marking tape; o The marking tape has special line on it which indicates the media within the autoclave machine has completed its process. o The line usually will turn into black stripes which indicated the completion of the processes, which in this case it will turn into black stripes after 20 minutes at 121C. o The marking tape is important in terms of determining the right amount of time and temperature used to boil or heated a subject.

Conclusion: - The media is successfully prepared within a petri dish