PCR Oct 9

Question 1

PCR

Answer 1

a rapid procedure for in vitro enzymatic amplification of a specific segment of DNA

Question 2

What are some applications for PCR?

Answer 2

Direct cloning from genomic DNA or cDNA
In vitro mutagenesis and engineering of DNA
Genetic fingerprinting of forensic samples
assays for presence of infectious agents
quantifying RNA transcripts
Covid19 testing

Question 3

DNA polymersase needs a ____ and will add nucelotide to the _____

Answer 3

primer, 3’OH end

Question 4

Forward primer, reverse primers

Answer 4

Forward - lower
Reverse - upper

Question 5

Basic PCR Protocol

Answer 5

1. Initial melting at 94 for 5mins
2. Denaturing of the DNA strands (94-95)
3. Annealing of oligonucleotide primers (55-65)
4. Extension with polymerase (72)
5. Repeat steps 2-4 for 25-30 cycles
6. Final extension at 72 for 5-10 mins

Question 6

When does the desired amplified product start to appear?

Answer 6

during the third cycle