ANALYSIS OF PLASMID AND GENOMIC DNA BY AGAROSE GEL Flashcards
What is a technique for separating and analyzing mixtures of charged molecules?
Electrophoresis
What is the term “to carry with electricity”.
Electrophoresis
T or F:
When placed in an electric field, these anion molecules migrate towards the anode for electrophoresis to occur?
True
What type of matrix used for electrophoresis?
agarose gel
T or F:
The larger the DNA fragment the harder it will be for it to move through the pores of the gel and the slower it will migrate.
TRUE
T or F:
Pore size is determined by the concentration of dissolved agarose.
TRUE
What is the role of the dyes that help monitor the progress of the gel?
they move in the same direction as nucleic acids during electrophoresis.
What is a DNA molecular weight marker?
a set of DNA fragments of known molecular sizes.
What are the type of electrodes used for agarose gel?
Black - Cathode (negative)
Red - Anode (positive)
Where does the TBE buffer go in the apparatus?
In the buffer Tank!
For the plasmid preparation, what is the purpose of adding the lysis solution after resuspending the cells?
Resuspension buffer is used for isolation of plasmid DNA by alkaline lysis method. The purpose of adding lysis solution is to disrupt exterior environment of a cell in a way that causes it to break open and release its components
Why do fragments of DNA in gel electrophoresis travel away from the wells?
DNA = negative charged, DNA will migrate towards the positively charged electrode
Explain how the percentage of agarose in a gel effects the gel electrophoresis.
it impacts the pore sizes (the higher the percentage of agarose, the smaller the pore size)
What is the purpose of the running buffer?
to create an electric field that allows for the movement of proteins through the gel during the electrophoresis process
What is the purpose of the tracking dye?
to visualize & track the movement of the DNA during electrophoresis experiments
