ANALYSIS OF PLASMID AND GENOMIC DNA BY AGAROSE GEL Flashcards

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1
Q

What is a technique for separating and analyzing mixtures of charged molecules?

A

Electrophoresis

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2
Q

What is the term “to carry with electricity”.

A

Electrophoresis

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3
Q

T or F:
When placed in an electric field, these anion molecules migrate towards the anode for electrophoresis to occur?

A

True

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4
Q

What type of matrix used for electrophoresis?

A

agarose gel

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5
Q

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.

A

TRUE

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6
Q

T or F:
Pore size is determined by the concentration of dissolved agarose.

A

TRUE

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7
Q

What is the role of the dyes that help monitor the progress of the gel?

A

they move in the same direction as nucleic acids during electrophoresis.

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8
Q

What is a DNA molecular weight marker?

A

a set of DNA fragments of known molecular sizes.

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9
Q

What are the type of electrodes used for agarose gel?

A

Black - Cathode (negative)
Red - Anode (positive)

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10
Q

Where does the TBE buffer go in the apparatus?

A

In the buffer Tank!

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11
Q

For the plasmid preparation, what is the purpose of adding the lysis solution after resuspending the cells?

A

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

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12
Q

Why do fragments of DNA in gel electrophoresis travel away from the wells?

A

DNA = negative charged, DNA will migrate towards the positively charged electrode

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13
Q

Explain how the percentage of agarose in a gel effects the gel electrophoresis.

A

it impacts the pore sizes (the higher the percentage of agarose, the smaller the pore size)

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14
Q

What is the purpose of the running buffer?

A

to create an electric field that allows for the movement of proteins through the gel during the electrophoresis process

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15
Q

What is the purpose of the tracking dye?

A

to visualize & track the movement of the DNA during electrophoresis experiments

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16
Q

What is the purpose of the standard marker (ladder)?

A

for size determination of dsDNA as well as PCR-generated DNA fragments

17
Q

What is the purpose of the gel green?

A

to stain DNA in agarose gel

18
Q

What is the purpose of the loading buffer in gel electrophoresis

A

To add density for sample loading

19
Q

What is the purpose of the wash solution in plasmid miniprep?

A

To remove the debris and impurities

20
Q

In which direction does DNA move during gel electrophoresis?

A

cathode -> anode
* because DNA is negatively charged therefore DNA will migrate towards the positively charged