DNA sequencing

Question 1

What is dideoxy chain termination also known as?

Answer 1

It is often called sanger sequencinge

Question 2

What is the DNA sequencing method developed to sequence?

Answer 2

Method developed to sequence DNA

Question 3

Robustness of DNA sequencing

Answer 3

Very robust with low error rate therefore reliable

Question 4

What are the steps involved in sequencing by dideoxy chain termination?

Answer 4

1. Generation of a template
2. Enzyming sequence reaction
   - DNA polymer makes multiple copies of DNA
3. Size separation of products by capillary electrophoresis
4. Detection of reaction product
5. Readout of sequence

Question 5

Sequencing by dideoxy chain termination is similar to PCR however what’s the difference?

Answer 5

• Only uses a single primer therefore linear amplification is not exponential
• Uses a DNA polymerase

Question 6

What does sequencing reaction involve?

Answer 6

Involves:

• Strand separation
• Annealing primer
• Extension
• Chain termination