Gene Analysis 2

Question 1

What techniques are used to study gene structure?

Answer 1

Restriction mapping
PCR
Southern analysis
DNA sequencing

Question 2

What does gel electrophoresis separate DNA fragments by?

Answer 2

Size

Question 3

Describe the process of restriction mapping

Answer 3

Cut with restriction enzymes

Run in electrophoresis

Question 4

Describe the process of polymerase chain reaction

Answer 4

DNA heated to 95C
A primer annealed at temp between 50-65C
Temp raised to 70C to allow thermostable DNA polymerase to synthesis 2 new strands

Question 5

What is southern blotting used for?

Answer 5

Allow specific fragments to be identified in a complex mixture

Question 6

What is the method of Southern blotting?

Answer 6

Preparation of restriction fragments
Gel electrophoresis 
Blotting onto a intracellular membrane
Hybridisation with radioactive probes
Probe detection

Question 7

What is DNA sequencing used for?

Answer 7

Determine order of bases in a strand of DNA in a 3’ to 5’ direction

Question 8

What is required for DNA sequencing?

Answer 8

Single stranded DNA template
Primer
dNTPs
ddNTPs
DNA polymerase

Question 9

What does cycle sequencing use?

Answer 9

DNA polymerase (themostable)
Single primer
Fluorescent ddNTPs