U4 LAB: ELECTROPHORESIS

Question 1

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

Answer 1

Electrophoresis

Question 2

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

Answer 2

solid, liquid

Question 3

2 main components of Electrophoresis chamber

Answer 3

• Chamber
• Power Source

Question 4

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

Answer 4

Chamber

Question 5

This supplies the electric current to the anode area

Answer 5

Power Source

Question 6

Positively charged pole, attracts negative particles

Answer 6

Anode (red)

Question 7

Negatively charged pole, attracts positive particles

Answer 7

Cathode (block)

Question 8

Flow of electric current

Answer 8

Cathode > Anode > Power Source

Question 9

Electrophoresis is used to separate?

Answer 9

negatively charged particles

Question 10

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

Answer 10

zwitterion

Question 11

Gel is commonly made up of?

Answer 11

Agarose

Question 12

Gel used in Protein Electrophoresis

Answer 12

Polyacrylamide gel

Question 13

The Electrophoresis run is dependent on the?

Answer 13

voltage used, and time

Question 14

DNA will be separated based on?

Answer 14

molecular weight

Question 15

Farther travels on gel

Answer 15

Lighter DNA

Question 16

Harder to travel

Answer 16

Heavier DNA

Question 17

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

Answer 17

Amphoteric

Question 18

Movement of buffer ions & solvent relatives to the fixed support

Answer 18

Electroendosmosis / Endosmosis

Question 19

Migration of small charged ions

Answer 19

Iontophoresis

Question 20

Five components of Electrophoresis

Answer 20

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

Question 21

Support Media

Answer 21

• cellulose acetate
• agarose gel
• polyacrylamide gel

Question 22

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

Answer 22

Ethidium Bromide (shaken then swirled)

Question 23

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

Answer 23

Tracking dye

Question 24

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

Answer 24

True

Question 25

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

Answer 25

False!

Question 26

Some manufacturers include what in their tracking dyes?

Answer 26

glycerol

Question 27

Common Tracking Dye

Answer 27

Cotton Phenol Blue

Question 28

Radioactive dye used to visualize DNA

Answer 28

Detection Dye

Question 29

Alternative fluorescent dye to EtBr and now often used

Answer 29

Gel Red

Question 30

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

Answer 30

Buffer

Question 31

Detection system

Answer 31

UV transilluminator

Question 32

Gel must be viewed in?

Answer 32

Gel Documentation Viewer

Question 33

Unit used in electrophoresis

Answer 33

Base pairs

Question 34

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

Answer 34

DNA ladder

Question 35

Left side of the gel contains?

Answer 35

DNA of known base pairs

Question 36

Increment of DNA ladder

Answer 36

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

Question 37

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

Answer 37

heavier Farther the sample, the lighter

Question 38

Electrophoresis can be used for?

Answer 38

quantification to see if there is a DNA fragment

Question 39

Why is Ethidium Bromide the best stain for DNA extraction?

Answer 39

intercalation or insertion between layers

Question 40

Amount of buffer needed is dependent on?

Answer 40

size of chamber

Question 41

Five Steps of Electrophoresis

Answer 41

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

Question 42

Fractions of Separated Proteins

Answer 42

1. Albumin 2. Alpha 1 3. Alpha 2 4. Beta 5. Gamma globulins

Question 43

Stains for Bands

Answer 43

- Amido Black - Ponceau S - Oil Red O - Sudan Black - Fat Red 7B - Coomassie Blue - Gold/Silver Stain

Question 44

Lower voltage for _____ periods of time

Answer 44

longer

Question 45

Higher voltage for _____ periods of time

Answer 45

shorter