PCR

Question 1

Basic PCR: Step 1

Answer 1

add primers and heat to separate strands

Question 2

Basic PCR: Step 2

Answer 2

cool to anneal primers

Question 3

Basic PCR: Step 3

Answer 3

Synthesis new DNA from the primers

Question 4

Basic PCR: Step 4

Answer 4

Using long strands as template, previous steps are repeated to synthesize short strands

Question 5

Basic PCR: Step 5

Answer 5

Short strands representing target sequence are repeatedly replicated

Question 6

Steps of RT-PCR

Answer 6

1) Total RNA extracted and mRNA separated using oligo (T) primer
2) Synthesis of cDNA by reverse transcription of mRNA using oligo (dT) to anchor primer.
3) Synthesis of second strand using oligo (dT) primer and primer in the presence of labelled dNTPs.
4) PCR amplification of cDNA using anchored oligo primer and a random primer.

Question 7

What does qPCR do?

Answer 7

amplify and detect changes in copy number

Question 8

What does RT-qPCR do?

Answer 8

combines Reverse Transcription and qPCR