Eukaryotic Lab

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Our Aim:
Our aim is to study the effects of adding a mutant to eukaryotic cells. The four parts of this experiment include finding the number of yeast cells provided, collecting RNA by using gel electrophoresis, synthesize cDNA, and determine the mutant by running a PCR.

Backgroung information: The budding yeast is an ideal experimental organism for genetic research. The yeast shares a common life cycle and cellular architecture with higher eukaryotes, and as a microorganism it is easily propagated and manipulated in the laboratory. The three types of mutants we are learning about in this lab are two that code for alpha and one that codes for beta-tubulin, which can have a few effects on the cell (Acosta 2015).
Materials and Methods:
Part …show more content…
Label your tube and prepare cDNA synthesis reaction in 0.2 ml PCR tube on ice. Add the specified samples to the tube. Mix gently and keep tubes on ice until putting them into the thermal cycler. When ready, load tubes in the thermal cycler for an hour and a half. When done store in the freezer for next lab.
Part IV – Polymerase Chain Reaction (PCR) for Mutant Strain Identification
Label the four tubes with your information and the name of the sample. Add each of the following to the tubes for a total of 50 uL reaction. Make sure the caps are on and load the tubes in the thermal cycler.
Gel electrophoresis of PCR samples:
Plug in the powerbase into an outlet using the adaptor plug. Remove the agarose gel from the package and remove comb from the cassette. Insert the gel starting from the right edge. Press firmly on the top and bottom and check for the red light, assuring the gel has but put in correctly. Load 20 uL of the PCR sample into each well and lastly, load 50 bp DNA ladder and fill any remaining wells with water. Place the amber filter on the chamber and select “E-Gel GP”, set the timer for 15 minutes and press the go button. When the time is up, press the red button, turn on the blue light and take a picture of
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