LAB PROTOCOLS
Overview
Barcoding Animal Life relies on a number of protocols for obtaining COI DNA sequences.

DNA Extraction
This stage involves breaking up tissues to make the DNA accessible for amplification. Different techniques are used depending on the quality of the individual tissue sample.

COI Amplification
Amplification (making copies) of the target DNA employs a technique called the Polymerase Chain Reaction (PCR). This is accomplished by placing the DNA extractions in a specialized chemical environment and cycling them through a specific temperature regime. A heat-activated DNA polymerase replicates the target DNA sequence with each cycle.

DNA Sequencing
At this stage, the products must be checked for both quantity and quality. Agarose gel electrophosesis is used to visualize the amount and size of DNA fragments present in the sample, and excess (non-target) DNA can be removed through gel-purification. Clean DNA is then prepared for sequencing by running a final PCR that incorporates dye terminator nucleotides.