Lab 6- DNA Barcoding: Polymerase Chain Reaction

purpose of PCR

to amplify the specific barcode fragment of DNA that we want to sequence

Temperature Change Throughout PCR (in order)

95 C→ 55 C → 72 C

PCR steps

Denaturation → Annealing → Synthesis

Why is the initial temperature set to 95 C

to denature enzymes

Taq polymerase purpose

it thrives in high temperatures so it doesnt denature during denaturation process

2 Primers needed during Annealing

Forward Primer and Reverse Primer

Forward Primer Direction

5’→3’

Reverse Primer Direction

3’→5’

What does Taq do during synthesis

begins to synthesize DNA from the two primers

What components go into the master mix

DNA polymerase, dNTPs, buffer, forward primer, reverse primer, water

what is dNTP

artificial nucleotides used to synthesize new DNA strand

What goes into Negative Control(N)

master mix

no DNA template

nuclease free water

what goes into Positive Control(P)

master mix

known bacteria

what goes into Sample(S)

master mix

diluted bacterial culture