DNA barcode usage plays a vital role since it is not linked with the dynamic or variable characteristics such as pattern or wing color that can be modified to the surrounding environment.
DNA barcode contains a section of DNA, inherited from the parent to the offspring. Independent mitochondrial DNA is very useful than relying on Morphology which sometimes may lead to the wrong decisions.
It allows the Scientist to collect the samples from a 50-year-old specimen because of its sustainability. DNA barcoding is less expensive and a great alternative to genome sequencing.
Barcoding contains many organisms of butterflies which helps the researchers to understand the biodiversity of life on the Earth’s environment.
Morphological Identification: The specimen is identified based on the morphological characteristics based on the wing pattern. But it is not sufficient for carrying the species identification. Thus to ensure the accurate identification of the specimen, it is been cross verified with Swiss butterfly taxonomy. DNA Sample: Butterflies tissue samples have to be collected that can be attained from a live butterfly which is preserved and processed. Sometimes, the samples from the existing collection are possible from the place such as the zoo and historical museum. A little of five mg is needed from these butterflies to carry DNA barcoding. Thus, the organism can be returned unharmed to the environment. Once the sample is collected, it is preserved in a safer place for further analysis. Genetic Analysis: DNA isolation is a method that has numerous toolkit that allows the rapid isolation of DNA from the butterfly tissue samples. In most cases, the physical and chemical process is used to break the cells and denature the cellular proteins. Thus, the obtained concentrated solution of cellular DNA is used as a PCR template. Polymerase chain reaction (PCR): The template strand (isolated from the tissue sample) is used to synthesize a new DNA strand by the usage of DNA polymerase. DNA template, DNA primers, and DNA polymerase are added to a buffered solution containing excess nucleotide (A, T, G, and C) along with the cofactors (eg.Mgcl) and later carried with alternating hot and cold cycles. DNA Sequencing: It is similar to that of the PCR technique which involves a small change in nucleotide to terminate the activity of the polymerase. Fluorescent dyes are used to label these nucleotides which makes them visible and can be detected by a camera. In this sequencing section, the DNA polymerase copies the template strand by chance, it will rarely insert the terminator nucleotide instead of a normal nucleotide. Thus, it terminates the process, and polymerase moves on to the next new strand creation. Therefore, the last copied DNA strand is labeled with the fluorescent dye. After the completion of this cycle, it is processed through the high percentage polyacrylamide gel (PAGE) and the process is recorded by the camera as it moves through the gel. The appropriate DNA sequence is been transferred and stored in a software program.
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