PCR- Micro 4130

Micro - Exam 1

PCR primers

DNA primers, become part of the product. Outline the product to be amplified and provide the 3’-OH to extend off of

Denaturing DNA with high heat

We use this method because there is no replisome/helicase to open the DNA

Steps of PCR

Denature template, anneal primers, elongate, repeat

Components of PCR

Template, polymerase, dNTPs, primers, heat, thermocycler

Designing primers

Start at very end of wanted sequence, because primers become part of the product! DNAP goes 5’-3’, so primers must go the same way. Upstream primer is identical to the sequence, and the downstream is complementary