PCR

What is polymerase chain reaction (PCR)

• Used to use bacterial cloning
  - Slow
  - Labor intensive

• Needed a faster way to copy a gene of interest

who was Dr. Kary Mullis

• Award Nobel Prize in 1993

• Revolutionary approach to rapidly amplifying specific fragments of DNA

• PCR enables vast quantities of specific DNA to be produced extremely quickly & at a low cost

What is polymerase chain reaction (PCR)

• Used to use bacterial cloning
  - Slow
  - Labor intensive

• Needed a faster way to copy a gene of interest

who was Dr. Kary Mullis

• Award Nobel Prize in 1993

• Revolutionary approach to rapidly amplifying specific fragments of DNA

• PCR enables vast quantities of specific DNA to be produced extremely quickly & at a low cost

Taq polymerase

• Isolated from Thermus aquaticus → ___________

• Functions best at 72ºC
  - Can survive up to 97.5ºC

Denaturation

95 celicus → creates ssDNA template

annealing

55-60 celicus → primers bind to the area around the target gene

extension (elongation)

72 celicus → tax builds off primers synthesizes target gene

PCR Applications

• monitor bacterial, fungal, & viral infections

  • COVID

• genotyping

  • ideas inheritance, forensic DNA analysis, paternity testing

• animal specific identification

  • trafficking

  • supplement ingredients

what is quantitative PCR

very sensitive, which means it can detect single copies of DNA

what are the different applications

• low copy number DNA

• Environmental DNA

low copy number DNA

ancient or very degraded (bones, ashes, poo)

environmental DNA (eDNA)

detect elusive or invasive species, pathogens