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MT6319 MOLBIO > U4 LAB: ELECTROPHORESIS > Flashcards

U4 LAB: ELECTROPHORESIS Flashcards

1
Q

Process of separating the charged constituents of a sample by means of an electrical current

A

Electrophoresis

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2
Q

Movement of charged particles through a _____ medium in a _______ buffer

A

solid, liquid

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3
Q

2 main components of Electrophoresis chamber

A
  • Chamber
  • Power Source
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4
Q

Tank-like, where solid media and liquid buffer is placed

A

Chamber

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5
Q

This supplies the electric current to the anode area

A

Power Source

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6
Q

Positively charged pole, attracts negative particles

A

Anode (red)

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7
Q

Negatively charged pole, attracts positive particles

A

Cathode (block)

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8
Q

Flow of electric current

A

Cathode > Anode > Power Source

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9
Q

Electrophoresis is used to separate?

A

negatively charged particles

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10
Q

Functional group molecule in which at least one has a positive electrical charge, based on isoelectric point

A

zwitterion

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11
Q

Gel is commonly made up of?

A

Agarose

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12
Q

Gel used in Protein Electrophoresis

A

Polyacrylamide gel

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13
Q

The Electrophoresis run is dependent on the?

A

voltage used, and time

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14
Q

DNA will be separated based on?

A

molecular weight

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15
Q

Farther travels on gel

A

Lighter DNA

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16
Q

Harder to travel

A

Heavier DNA

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17
Q

Has a net charge that can be either positive or negative depending on pH conditions

A

Amphoteric

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18
Q

Movement of buffer ions & solvent relatives to the fixed support

A

Electroendosmosis / Endosmosis

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19
Q

Migration of small charged ions

A

Iontophoresis

20
Q

Five components of Electrophoresis

A
  1. driving force
  2. support medium
  3. buffer
  4. sample
  5. detecting system
21
Q

Support Media

A
  • cellulose acetate
  • agarose gel
  • polyacrylamide gel
22
Q

When utilizing electrophoresis for DNA observation, what stain must be used?

A

Ethidium Bromide (shaken then swirled)

23
Q

This is added in order to track the migration of analyte across the gel.

A

Tracking dye

24
Q

T/F: The tracking dye migrates ahead of the bond or the electrolyte of interest that you are trying to separate.

A

True

25
Q

T/F: The electrophoretic run can continue even if the tracking dye nears the end of the gel.

A

False!

26
Q

Some manufacturers include what in their tracking dyes?

A

glycerol

27
Q

Common Tracking Dye

A

Cotton Phenol Blue

28
Q

Radioactive dye used to visualize DNA

A

Detection Dye

29
Q

Alternative fluorescent dye to EtBr and now often used

A

Gel Red

30
Q

This will give proteins the net electric (negative) charge that allows them to move from cathode to anode.

A

Buffer

31
Q

Detection system

A

UV transilluminator

32
Q

Gel must be viewed in?

A

Gel Documentation Viewer

33
Q

Unit used in electrophoresis

A

Base pairs

34
Q

This will serve as reference to know how many base pairs your DNA is.

A

DNA ladder

35
Q

Left side of the gel contains?

A

DNA of known base pairs

36
Q

Increment of DNA ladder

A

50 (Start from 50, 100, 150, 200 and so on)

37
Q

The closer the sample is to the well, the _____ it is

A

heavier

Farther the sample, the lighter

38
Q

Electrophoresis can be used for?

A

quantification to see if there is a DNA fragment

39
Q

Why is Ethidium Bromide the best stain for DNA extraction?

A

intercalation or insertion between layers

40
Q

Amount of buffer needed is dependent on?

A

size of chamber

41
Q

Five Steps of Electrophoresis

A
  1. Loading of patient sample
  2. Electrophoretic migration
  3. Wash and fix
  4. Staining
  5. Visualization (Qualitative)
  6. Quantification
42
Q

Fractions of Separated Proteins

A
  1. Albumin
  2. Alpha 1
  3. Alpha 2
  4. Beta
  5. Gamma globulins
43
Q

Stains for Bands

A
  • Amido Black
  • Ponceau S
  • Oil Red O
  • Sudan Black
  • Fat Red 7B
  • Coomassie Blue
  • Gold/Silver Stain
44
Q

Lower voltage for _____ periods of time

A

longer

45
Q

Higher voltage for _____ periods of time

A

shorter

MT6319 MOLBIO (10 decks)

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