How do you isolate and copy the COI gene from your salmon sample?
Once you have obtained your salmon sample from a local store, you need to isolate the DNA from it and make many copies of the COI gene using the polymerase chain reaction (PCR).
Place the following steps of the DNA extraction and PCR process in the correct order. Not all labels will be used.

1. Disrupt cell, nuclear, and organelle membranes to release their contents into solution.
2. Add Tag, primers, and dNTPs to muscle tissue.
3. Cut out a clean section of muscle tissue.
4. Add Tag, primers, and dNTPs to the DNA sample.
5. Use a thermal cycler to denature, anneal, and extend the target sequence (the COI gene) in 35 cycles.
6. Eliminate non-DNA molecules using wash buffers.

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Explanation: Polymerase Chain Reaction or PCR is a lab technique used to make numerous copies of determined section of a DNA. The process requires 5 ingredients to perform:

1) The DNA template to be copied, which can be obtained by disrupting the nuclear membrane of a cell and releasing its components into a solution;

2) Short sequences of DNA, called primers, designed to be complementary to the DNA to be copied;

3) DNA nucleotide bases, also known as dNTPs (A, T, C and G);

4) Taq polymerase enzyme;

5) Buffer to ensure optimal conditions to the reaction;

PCR involves three stages:

Denaturing: this stage takes 15 to 30 seconds and consists of putting the DNA and the other ingredients in a thermal cycler, heated to 94-95°C. In that temperature, the DNA to break into two strands, in a process called Denaturation;Annealing: the temperature is reduced to 50-65°C, so the primers can attached itself to a specific location on the stranded DNA. This step initiates the synthesis, because the polymerase enzyme can only add bases to a double strand of DNA. Once bounded, it takes 10 to 30 seconds to make a new complimentary strand of DNA from the model;Extending: In this stage, the temperature is increased to 72°C, which enables the Taq polymerase enzyme. This enzyme comes from a bacteria, which supports high temperatures, and has a role of builiding the complimentary strand by binding the primer and adding the DNA bases to the single strand. This creates a new molecule of DNA. The time this step takes depends on the length of the DNA to be copied;

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