Polymerase Chain Reaction - 16A

What is the purpose of PCR?

To replicate specific segments of DNA

What is the name of the 2 DNA primers?

Bact 8F and 1530 R

What are the ingredients for PCR?

• dNTP’s

• Taq polymerase

• 2 DNA primers

• DNA Template (chromosomal DNA extracted from the unknown)

What is Taq Polymerase derived from?

Thermus aquaticus which allows DNA not to get inactivated from heat and can build phosphodiester bonds at 72 degrees celsius

What are the 3 steps of PCR?

Denaturation, Annealing, and Extension

Explain step 1 of PCR.

Denaturation - 94/95 degrees celsius heat breaks the H bonds apart making single-stranded DNA

Explain step 2 of PCR.

Annealing - cooling the temperature down to 55 degrees Celsius so hydrogen bonds can form between the nucleotides dNTP, and primers

Explain step 3 of PCR.

Extension - heated to an optimum temperature of 72 degrees celsius allowing Taq polymerase to form phosphodiester bonds b/w the growing strand of DNA and new dNTPs