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Lab 4: Using the Spectrophotometer to Measure the Rate of Bacterial Cell Division: A Bacterial Population Growth Curve

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Lab 4:  Using the Spectrophotometer to Measure the Rate of Bacterial Cell Division: A Bacterial Population Growth Curve (Photo:Miamioh) Introduction     The purpose of this lab will ultimately be to observe the growth of an Escherichia coli population in its lag phase, its exponential growth phase and its stationary phase.  Each group will measure the turbidity of different E. coli samples all taken at different times to get a wide range of measurements that will then construct a growth graph.  The biggest issue will most likely be the counting of the hemocytometerns that contain the E. coli .     While it is different for every bacteria, E. coli has a doubling time as short as 20 minutes. However, once they are placed in a growth medium (in this case LB broth), they will take some getting used to their environment. This period of adjustment is called the lag phase and will seen as the initial flatline on the growth curve graph.  Once th...
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Lab 6: Coliform Determination by the Membrane Filter Technique

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Lab 6:  Lab Coliform Determination by the Membrane Filter Technique (Photo: Steve Nease) Introduction     The purpose of this lab will ultimately be to familiarize ourselves with counting coliform microorganism colonies as well as another plating method called the membrane filter technique.     The total viable count is based upon the ability of a microorganism placed in a nutrient environment to grow.  While individual microorganisms cannot be seen with the naked eye, they can grow in colonies which are visible to any plain Jane.  Counting the colonies gives an indication of the number of viable colony forming units (CFU) present in the original sample.  However, the appropriate growth medium must be chosen first to get to the counting phase.  This part is crucial because the accuracy will be best if the chosen medium is as similar as possible to the sample environment.     The membrane filter technique uses the semi pe...
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Lab 8: DNA Isolation

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Lab 8:  DNA Isolation (Photo: HSHP Biology) Introduction     The purpose of this lab was ultimately to familiarize ourselves with the PowerSoil DNA Isolation Kit to successfully purify a DNA sample from a pure E. coli culture in LB broth.. The first step will be obtaining the cells, then through use of the kit and instructions we will break down the membrane and release the DNA from the cells. The next step will be to precipitate the DNA and finally, purify the sample. "The PowerSoil DNA is intended for use with environmental samples containing a high humid acid content including difficult soil types such as compost, sediment and manure...The isolated DNA has a high level of purity allowing for more successful PCR amplification of organisms from the sample." (MoBio) Since the sample we are using is already a pure E. coli sample, all of the purification is a tad overkill but will be done anyways according to the provided instructions. Another important thing to no...
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