CULTURING OF MICROBS

CULTURING OF MICROBS -...

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http://www.bioreactors-medorex.com/index.html Date: 2-7-2007 The first practical approach to the whole culturing process. At 9’o clock, our group members which consists of two people ( Chai Kang Han and Loh Hong Siang ) meet at the engineering staff office. After instruction and final discussion with Dr. Marwan, we went to the bio-medical laboratory. Here, the first time we approach everything with our own hands. After re-calculating the ingredients for the LB mixture which is also known as “rich media” which is suitable for all microbs in the atmosphere to grow in it. Thus, we do not want this to occur, as this will contaminate our culture and distort our readings. Precautions are taken to prevent this from happening. The precautions taken are as below:- 1. Sterilization of the apparatus. Sterilization is a very crucial step in bio- technology. This is our first priority for the whole culturing course. There are 3 ways to sterilize equipment and culture media. They are autoclave which sterilize utilizing steam at pressure and high temperature. The second way is flaming the nozzles of bottles or inlet to kill microbs. The third way is to use ultra-violet light to sterilize but direct eye contact is harmful. Ethanol at concentration of 70% can also be a sterilizing agent. This is use to sterilize some of the equipments before it is send to the fume hood. 2. The prepared LB media is sterilized with some measuring cylinders and tips for the micro-pipettes. This is to prevent contamination from any unwanted microbes from the air. After some time, the autoclaved LB media is left inside the fume hood to cool to atmospheric temperature. However, the media with agar in it is left to stir on the magnetic stirrer as its temperature decreases. A better method to allow the bottle to cool down in shorter time is to let it run under tap water and slowly spins the bottle. Care as the bottle is hot and it is more brittle if it hits the sides of the sink it will crack. At 5’o clock in the afternoon, one of the in-charge lecturer Mr. Rashidan inoculate two samples from the original storage source of the bacteria into the prepared rich-media solution to culture the microbs from the freezer which is believed to yet be in their dormant state. The specimen inoculated are “BSM Klang 2” and “BSM Klang 3a”. After cleaning up the apparatus and autoclaving some of the apparatus such as the tips for the micro-pipettes and measuring cylinders, we left the fume hood turned on as instruction from the lab assistant Mr. Khairul, we left the laboratory. 1
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Date: 3-7-2007 At 8.30 a.m., we enter the lab with our lab coats and started the experiment for the day. The fume hood was turn on and u.v. rays are allowed to shine onto the fume hood for 15 minutes before the exhaust fan of the hood is turned on. After which 70% ethanol is used to cleaned the base of the fume hood where the sample is being taken is cleaned. Observations shows the agar plates we prepared yesterday was solidify over night as
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This note was uploaded on 06/06/2011 for the course CHEM 3040 taught by Professor Reddy during the Spring '10 term at Taylor's.

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CULTURING OF MICROBS -...

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