lecture 2 - amplifying DNA

Question 1

components of PCR

Answer 1

template (ds DNA)
2 primers (small ssDNA)
polymerase
dNTPs (deoxyribonucleotide triphosphates)
magnesium (co factor for DNA polymerase enzyme)
buffer (maintains pH)

Question 2

C-G 3 hydrogen bonds

Answer 2

A-T 2 hydrogen bonds

Question 3

hydrogen bonds are very

Answer 3

weak
when they’re combined they become strong

Question 4

DNA always read it

Answer 4

5’ to 3’

Question 5

lagging strand

Answer 5

primer copies in sections (okazaki fragments)
ligase joins these fragments together

Question 6

replication in eukaryotic cells

Answer 6

primase - create RNA and bind to to the DNA (create primers)
alpha and delta polymerase work on lagging strand and epsilon wonks of leading strand
nucleases - remove RNA primers and ligase then joins the fragments together
ssDNA binding proteins - binds to ssDNA to protect it
sliding clamps - attach to polymerase and stop it falling off

Question 7

pyrimidines

Answer 7

cytosine and thymine

Question 8

purines

Answer 8

adenine and guanine

Question 9

taq polymerase

Answer 9

found in hot springs

Question 10

melting temp of primers

Answer 10

50 - 60 degrees
the primer pairs mp should be within 5 degrees of eachother
primer pairs shouldnt have a complementary region

Question 11

magnesium

Answer 11

a co factor
acts to enhance the enzymic activity (specifically of DNA polymerase) thereby supporting DNA application
magnesium binds into active site of polymerase catalysing the reaction

Question 12

buffer

Answer 12

tris HCL

Question 13

copies =

Answer 13

2^n (n = cycles)