| Lab Techniques |
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Nic Theobald | Yilectronics |
| Introduction: Most effort in ensuring good results and repeatability is as simple as being consistent while using tools and methods. Here are a few brief descriptions and videos on the many techniques and methods used in these tutorials. Also included are some images from when I participated in this Lab... Aseptic Technique/Bunsen Burners/Inoculation/: Aseptic technique is the practice of preventing contamination. This could be as quick and simple as wiping down the lab bench or as lengthy as autoclaving your tools. It is extremely important to plan and organize your work station according to the work you will be performing. For example, it is bad practice to unnecessarily expose pipette tips. When transferring liquids, you should make sure that the receiver, source, and pipette tip tray are ready so that the all three are exposed for the least amount of time possible. This will be discussed in more detail as it becomes more relevant. This video covers the operation of a Bunsen burner, how to handle sterile equipment, and how to inoculate plate and liquid cultures. It also shows how to inoculate a stab culture, but we won't be using that method... Before handling sterile equipment that will directly or indirectly touch the bacteria you should wipe down your work space, wash your hands, and put on gloves. This helps mitigate the risk of contamination. Our lab didn't flame the mouth of the test tubes, so I will update this page when I get more information. Take special note of how the test tubes are handled (the cap is never set down and is only off when absolutely necessary). Same goes for containers, only take the cover off when necessary and never set it down. Cell culturing is the process of growing a large amount of cells under controlled conditions. In this lab, cell culturing is used to grow individual, distinct colonies of cells that can be directly used later. This lab started with precultured liquid and plate media which were both used to inoculate our own culture plates. The culture plates were then incubated for approximately 48 hours. Two colonies from one of the culture plates were used perform PCR. Note: This
video shows how to inoculate plate media without creating a gradient of
bacteria colonies. The lab procedures will show how to properly segment a plate
culture and inoculate in a manner that will create a gradient...
Pipetting: Pipetting is the process of accurately measuring and dispensing small volumes of liquid. This tutorial uses micro scale and milli scale pipettes. Plate Streaking (general overview): Plate streaking is the process of applying cells to a media plate in a manner that creates a gradient of distributed cells. The goal is to create a gradient from a large mass of cell growth down to a few small colonies. These small colonies make it significantly easier to isolate a particular colony for further use. |