PCR

Question 1

Who developed PCR?

Answer 1

Kary Mullis

Question 2

Give the equation to count the total number of DNA after PCR cycle.

Answer 2

Nt = No x 2^n

Question 3

Name the divalent cation that helps in coactivating polymerase

Answer 3

Mg2+ or Mn2+

Question 4

Name the other DNA polymerase that has proof reading activity

Answer 4

Thermus thermophyllus, Pyrococcus furiosa, Pyrococcus woesei and Thermococcus litoralis

Question 5

Name the three steps of PCR and their temperature

Answer 5

Denaturation - 90 to 100°C
Annealing - 40 to 60°C
Extension or DNA synthase - 70°C

Question 6

What is nested PCR

Answer 6

PCR that uses two sets of primer to increase the specificity of DNA amplification and reducing non specific DNA amplification

Question 7

Name the dyes used in qPCR

Answer 7

SYBR green and TaqMan Probe

Question 8

What is RT PCR?

Answer 8

The PCR which converts RNA to cDNA

Question 9

Name the microorganism for reverse transcriptase

Answer 9

Thermus thermophyllus

Question 10

Thermus thermophyllus needs the presence of what to work?

Answer 10

MnCl2

Question 11

What is inverse PCR

Answer 11

The amplification of unknown DNA without using any primers