Part 4: Polymerase Chain Reaction
We will dilute the DNA that you isolated from your salmon sample, and pipette a small amount of the dilution into a PCR tube along with a premade "PCR master mix." The master mix contains fish-specific COI primers to amplify part of the mitochondrial cytochrome-c oxidase gene as well as Taq polymerase, deoxynucleotides (dNTPs), and buffer for optimal salt & pH conditions. The DNA is diluted to reduce the chance that contaminants (e.g., polysaccharides or proteins) not fully washed away during DNA isolation may inhibit the reaction, which might cause the PCR reaction to fail.
Your PCR tubes will then be placed into a thermal cycler and will undergo 40 cycles of amplification. After the PCR had completed, your tube will be held for you until next week, where you will use gel electrophoresis to check your PCR product before sending it off for sequencing.
WEAR GLOVES while setting up your PCR reaction!