Electrophoresis Flashcards

1
Q

What does electrophoresis allow us to do ?

A

Separate DNA fragments according to length

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2
Q

What is the electrophoresis separation technique based on ?

A

The movement of charged molecules in an electric field

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3
Q

What does DNA consist of ?

A

Two sugar phosphate backbones joined by complementary base pairs

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4
Q

What are phosphates electric charge ?

A

Negative

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5
Q

Why is DNA attracted to the positive electrode -anode ?

A

Phosphate backbone is negative

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6
Q

Why are all DNA fragments attracted equally to the anode ?

A

The charge to mass ratio is uniform throughout its length

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7
Q

What is a gel matrix ?

A

A cross linked substance that acts as a molecular sieve

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8
Q

What gel matrix is used in DNA electrophoresis ?

A

A polysaccharide called agarose

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9
Q

What is the purpose of agarose ?

A

Impedes the migration of DNA molecules

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10
Q

What size molecules find it more difficult to travel through the matrix ?

A

Larger

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11
Q

What size molecules travel further ?

A

Smaller

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12
Q

How is the gel matrix made ?

A

Agarose powder is dissolved in a buffer with a high ion concentration

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13
Q

What is the function of the wells in the matrix ?

A

Keeps the DNA samples apart

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14
Q

Where is the gel matrix placed ?

A

Submerged in a high salt buffer

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15
Q

What results in an electrical conductor ?

A

The high ion concentration in the buffer and the gel itself

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16
Q

What is the purpose of glycerol in loading buffer ?

A

To make samples dense so they sink to the bottom of wells

17
Q

What allows us to see what is happening in the matrix ?

A

Blue stain - Bromo-phenol blue

18
Q

What type of light is the gel viewed under ?

A

UV

19
Q

What does each band of DNA consist of ?

A

DNA of a particular length

20
Q

What is the DNA ladder ?

A

The lane containing the length standards

21
Q

What is the length of DNA molecules written in terms of ?

A

The number of base pairs

22
Q

What is a restriction digest ?

A

Where DNA is cut into specific fragments

23
Q

What is gel electrophoresis useful for ?

A
  1. Checking restriction digests
  2. Checking PCR products
  3. Isolating a desired DNA fragment
24
Q

What is a restriction enzyme ?

A

Cuts DNA at a specific base sequence to give a mixture of fragments known as a digest

25
Q

Is DNA positively of negatively charged ?

A

Negatively