Molecular Diagnostics Detection of Nucleic Acids: Electrophoresis

Question 1

Terminology:
Movement of molecules by electric current

Answer 1

Electrophoresis

Question 2

TRUE or FALSE:
Anions travel toward the cathode because it has a positive charge

Answer 2

FALSE - the anode has a positive charge

Question 3

TRUE or FALSE:
Cations, positively charge particles, will travel toward the cathode because it has a negative charge.

Answer 3

TRUE

Question 4

This method of testing is often used for separating, identifying, and purifying DNA fragments

Answer 4

Electrophoresis

Question 5

pH range at which DNA is negatively charged

Answer 5

7.0-8.0

Question 6

Smaller DNA pieces migrate (faster/slower) than larger DNA pieces

Answer 6

Faster

Question 7

4 Types of electrophoresis gel systems

Answer 7

-Agarose Gel Electrophoresis
-Pulsed Field Gel Electrophoresis (PFGE)
-Polyacrylamide Gel Electrophoresis (PAGE)
-Capillary Electrophoresis

Question 8

Terminology:
Polysaccharide polymer from seaweed

Answer 8

Agarose

Question 9

Agarose Gel Electrophoresis can be used for nucleic acids ranging from ____ base pairs to _______ base pairs:
a- 50,000 - >250,000
b- 5 - 500
c- 200 - 50,000

Answer 9

c- 200-50,000 base pairs

Question 10

Pulsed-Field Gel Electrophoresis can be used for nucleic acids ranging from ____ base pairs to _____ base pairs:
a- 50,000 - >250,000
b- 5 - 500
c- 200 - 50,000

Answer 10

a- 50,000 - >250,000

Question 11

TRUE or FALSE:
Agarose Gel Electrophoresis is the most commonly used gel system in clinical labs

Answer 11

TRUE

Question 12

Which gel electrophoresis method uses alternating electrical fields?

Answer 12

Pulsed-Field Gel Electrophoresis

Question 13

This gel electrophoresis method is used for bacterial typing for epidemiology

Answer 13

Pulsed-Field Gel Electrophoresis

Question 14

This gel electrophoresis method uses a synthetic substance which allows for more controlled and uniform pores.

Answer 14

Polyacrylamide Gel Electrophoresis

Question 15

TRUE or FALSE:
Polyacrylamide is dangerous in powdered form because it is a neurotoxin

Answer 15

TRUE

Question 16

Polyacrylamide Gel Electrophoresis can be used for nucleic acids ranging from ____ base pairs to _____ base pairs:
a- 50,000 - >250,000
b- 5 - 500
c- 200 - 50,000

Answer 16

b- 5 - 500

Question 17

This gel electrophoresis method is used for nucleic acid sequencing, mutation analysis, and research

Answer 17

Polyacrylamide Gel Electrophoresis

Question 18

Gel electrophoresis method that uses a small bore capillary tube to separate particles by size and charge

Answer 18

Capillary Electrophoresis

Question 19

Gel electrophoresis method used in forensic application, parentage testing, and DNA sequencing

Answer 19

Capillary Electrophoresis

Question 20

What are the 2 most common buffers used in gel electrophoresis?

Answer 20

-TBE (Tris borate EDTA)
-TAE (Tris acetate EDTA)

Question 21

In which buffer will DNA migrate twice as fast?

Answer 21

Tris acetate EDTA (TAE)

Question 22

What could happen if the buffer is too concentrated?

Answer 22

Increased conductivity creating more heat which could melt the gel or denature the sample

Question 23

What are 3 methods of visualizing nucleic acids?

Answer 23

-Ethidium bromide
-Silver Stain
-Fast Blast DNA stain

Question 24

Which visualization method is known to be a carcinogen?

Answer 24

Ethidium bromide

Question 25

Which visualization method is prone to overdevelopment?

Answer 25

Silver stain

Question 26

Stain used for protein visualization

Answer 26

Silver stain

Question 27

Dye that intercalates between bases if dsDNA

Answer 27

Ethidium bromide

Question 28

Advantage(s) of which gel electrophoresis method?: -Large range of separation -Easy to set up and perform

Answer 28

Agarose Gel Electrophoresis

Question 29

The purpose of ________ is to carry current and protect the samples during electrophoresis

Answer 29

Buffer