11 Jun Demonstrate the processes of DNA Electrophoresis
Question
Demonstrate the processes of DNA Electrophoresis and PCR
Approximately 1 to 11/2 hours to view videos and answer questions
Materials: none
Procedure
DNA Electrophoresis
Watch each of the following videos TWICE before answering the questions below: How to Use a Pipette (https://www.youtube.com/watch?
v=mUikNHvtBDE&feature=youtu.be) (5:11 minutes)
How to make and run an agarose gel (DNA Electrophoresis) (https://www.youtube.com/watch?v=EOu6hHqofuQ&feature=youtu.be ) (4:54 minutes) Questions
1.
2.
3.
4.
5.
6. What is agarose and why is it used to separate DNA?
Why would a scientist use a higher versus a lower percent agarose solution when preparing a gel to separate DNA?
Electrodes are applied to facilitate the separation of DNA in the gel, what type of charge will be applied to the DNA?
How do you determine when the DNA has run far enough through the gel?
How would a scientist view the DNA on the gel, once it has been separated through gel electrophoresis?
A ladder is typically used to determine the size of DNA samples separated through gel electrophoresis. Would you expect shorter or longer fragments of DNA to run further on a given gel? Polymerase Chain Reaction (PCR)
Watch each of the following videos TWICE before answering the questions below: Polymerase Chain Reaction (PCR) (https://www.youtube.com/watch?
v=2KoLnIwoZKU&feature=relmfu (1:28 minutes)
Polymerase Chain Reaction (PCR) (https://www.youtube.com/watch?
v=JRAA4C2OPwg )(2:10 minutes)
Questions 1. When preparing a PCR reaction for the thermocycler, a researcher will add taq polymerase, DNA nucleotides, and a 5’ and 3′ primer, among other components. For each of the components listed here, please indicate their purpose in the reaction.
2.
What does PCR stand for and what is the general purpose?
3.
PCR is performed in a Thermocycler, where varying temperatures are applied for a number of cycles indicated by the researcher. Please list the temperatures mentioned in the video and the reason that each temperature is applied to the sample.
4.
When preparing PCR reactions, researchers often include a water control that contains all
of the components except the DNA. Why might a researcher want to include this sample?
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