PCR

Question 1

Steps to denaturing

Answer 1

Two strands of DNA are separated by heating DNA up to 96’C to weaken hydrogen bonds

Question 2

Steps to Annealing

Answer 2

• DNA is cooled to 55-65’C
• A primer is attached to each strand at the 5’ end and 3’ end
• Th primer is a section of DNA with a known sequence which attaches to a specific part of the single DNA chain

Question 3

Steps to elongation

Answer 3

• Increase heat up to 73’C in the Thermocycler
• Taq polymerase adds complementary nucleotides to the strand of DNA until it reaches the primer, which prevents from any more nucleotides being added

Question 4

What does repeating the steps of PCR 30-40 times do

Answer 4

• Original DNA sample is limited to you need lots for Gel electrophoresis
• Repeating the steps creates billions of copies of the DNA

Question 5

What do you need to do before PCR

Answer 5

• identify segment of DNA or desired chromosome or gene

- Make a copy of that segment