AGE

Question 1

To avoid overheating, take note of the

Answer 1

Voltage
Ionic strength

Question 2

Increased voltage…

Answer 2

Increased heat

Degradation of gel
Bad resolution

Question 3

Increased ionic strength?

Answer 3

Increased conductivity

Increased heat

Question 4

Migration pattern of DNA

Answer 4

Cathode to anode

Question 5

TYPES OF ELECTROPHORESIS

Answer 5

AGE
PAGE

Question 6

Most widely used
Larger range of separation
Low resolution

Answer 6

Agarose gel electrophoresis (AGE)

Question 7

AGE

Range of Concentration:

Answer 7

0.5%-2%

Question 8

AGE

Advantage:
Disadvantage:

Answer 8

It can separate large molecules like NA or DNA

Low resulting power

Question 9

High resolution
Suitable for smaller molecules

Difficult to make and expensive

Answer 9

Polyacrylamide gel electrophoresis (PAGE)

Question 10

PAGE

Range of Concentration:

Answer 10

3.5%-20%

Question 11

PAGE

Advantage:
Disadvantage:

Answer 11

High Resolution

More cumbersome - difficult to make and it’s more expensive than AGE

Question 12

Starch - medium

Cheapest
Poor resolution

Answer 12

Starch gel electrophoresis

Question 13

Agarose vs Polyacrylamide

Horizontal or vertical?

Answer 13

Horizontal
Vertical

Question 14

Agarose vs Polyacrylamide

Big gaps or small gaps in gel?

Answer 14

Bigger gaps that with
polyacrylamide

Smaller gaps

Question 15

Agarose vs Polyacrylamide

Separates large molecules or small ones?

Answer 15

Both, but better for large molecules
Both, but better for small molecules

Question 16

Agarose vs Polyacrylamide

Lets you tell small differences between sizes of bands?

Answer 16

No
Yes

Question 17

Agarose vs Polyacrylamide

Type of molecules run in this type of gel

Answer 17

Mostly DNA

DNA or protein

Question 18

The spaces in between the gel particles in the well are the_____.

It is where the sample will pass

Answer 18

pores

Question 19

We put the sample in the____.
• Upon applying electric current, the DNA molecule will migrate slowly

Answer 19

well

Question 20

Band

• Intact =
• Smudge =

Answer 20

good quality DNA

degraded DNA

Question 21

Highly purified uncharged polysaccharide derived from agar

Answer 21

AGAROSE

Question 22

Agarose

Dissolves when added to boiling liquid
When temperature lowers to about____, it gels/ solidifies

Answer 22

40°C

Question 23

• The lower the concentration, the____ the pores
Good separation for…

• The higher the concentration, the____ the pores
Good separation for…

Answer 23

bigger; large DNA fragments

smaller; smaller DNA fragments

Question 24

Held together by the formation of weak hydrogen and hydrophilic bonds (hydrocolloids)

Answer 24

Agarose gel

Question 25

BUFFERS FOR ELECTROPHORESIS

Answer 25

Tris-Acetate EDTA (TAE) Tris-Borate EDTA (TBE)

Question 26

Common running buffer for agarose

Answer 26

Tris-Acetate EDTA (TAE)

Question 27

TAE Working solution: Stock solution:

Answer 27

0.5 - 1.0 x 50 x

Question 28

Stronger buffering capacity Can be used for agarose or PAGE

Answer 28

Tris-Borate EDTA (TBE)

Question 29

TBE Working solution: Stock solution:

Answer 29

0.25 x 5 x

Question 30

TAE vs TBE Difference in components

Answer 30

Glacial acetic acid Boric acid

Question 31

Intercalates DNA to its planar structure

Answer 31

ETHIDIUM BROMIDE (EtBr)

Question 32

EtBr UV absorbed by DNA at_____ is transmitted to the dye

Answer 32

160 nm

Question 33

EtBr in UV light emits a ____color (fluorescence) at_____ detected by the naked eye

Answer 33

red-orange ; 590 nm

Question 34

SAFER ALTERNATIVES TO ETHIDIUM BROMIDE (EtBr) EtBr is carcinogenic and toxic

Answer 34

SYBR® Safe DNA Gel Stain Gel Red®

Question 35

is to determine the size of the molecule Our reference on the estimation of the amount of base pairs our sample has or the size of our sample

Answer 35

Ladder