Module 6: Manipulating Genomes

Question 1

Principles of DNA sequencing

Answer 1

1. Primers added to the template DNA (start of gene) to be sequenced
2. DNA polymerase, nucleotides and dideoxynucleotides are added to the template strand
3. DNA polymerase makes many copies of the template whenever a ddNTP is incorporated the sequence terminates.

Question 2

Principles of Polymerase chain reaction

Answer 2

1. Thermocycler heats mixture to 94ºC to separate strands
2. Mixture is then cooled to 68ºC to allow primers to bind to template
3. Mixture is then heated to 72ºC to allow tax polymerase to add nucleotides to the new strand of DNA