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Nt1310 Lab 1

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1. What is the purpose of flaming the loop before and after use?

This process sterilizes the inoculating loop before the experiment. By passing the inoculating loop through the warmest part of the flame (the tip of the inner blue flame), the heat kills any microbes and prevents contaminating microorganisms from interfering with the integrity and accuracy and contaminating your experiment. This process is known as flame sterilization.

Flaming the loop after the experiment once again kills any contaminating microorganisms (bacteria) that may still be on the loop from the previous colony/culture/broth/medium it came into contact with. This prevents the spread (cross contamination) of unwanted/possibly contaminating bacteria which may contaminate the experiment and yield incorrect/unreliable results.

2. …show more content…
Why must the loop be cooled before touching it to a culture? How do you know when it is cool?

If the inoculating loop is still hot/has sufficient heat, it may destroy the bacteria it comes into contact with in the culture. This may occur as the inoculating loop is exposed to extreme heat during flame sterilization. As a result, one would not be able to, for instance, successfully transfer bacteria to another medium as the bacteria would be killed by the heat and thus not grow in the new medium. Allow a few seconds for the loop to kill thereby avoiding killing the bacteria.

Wait for a few seconds after flame sterilization. One may gently tap the loop on a clear area on the agar plate (where there is no growth of bacteria) in order to test if it is sufficiently cool. If the agar is damaged/one sees evidence of a heated loop, it is still too hot. It is crucial to avoid breaking/damaging the agar surface.

Do not place the inoculating loop on a surface/desk to cool after flame sterilization as this may once again contaminate the loop and defy the whole purpose of flame sterilization.

3. Why should the lids of the tubes not be placed on the

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