First off, start by wiping down your work surface and your general area with some tissue soaked in ethanol. Once the surfaces have been cleaned, set up a bunsen burner and light it, and open the air hole fully. This will cause a vacuum of air and keep bacteria away from your experiment. It can also be used to sterilise tools e.g. innoculating loops. Once this has been done, you are ready to begin applying your bacterium to your agar and create your culture. In order to move the bacterium from the test tube to the dish, however, many precautions must be taken. You must take the lid off the test tube near to the flame, and run the neck of the tube through the flame to kill any unwanted bacteria. It is also advised to hold the lid between your palm and little finger in order to avoid contamination. Once you have done this, simply dip the loop into the solution of bacteria and remove it again. Pass the neck through the flame again to sterilise it, then replace the lid and put it to one side. Once you have some bacteria on the loop, spread it evenly on the dish by brushing the loop on the surface. to ensure it is evenly distributed, rotate it 90 degrees four times, and between each turn brush the loop on the agar. Ensure you keep the lid low above the dish to shield it from bacteria. Once you have spread it evenly, simply replace the lid and write your initials and the date around the edge of the base of the dish. You should also tape the lid to the base to prevent bacteria getting in due to the lid falling off.
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