WHAT IS DNA BARCODING? | Online Biotech Notes

Online Biotech Notes

WHAT IS DNA BARCODING?

• DNA Barcoding is a method of identification of species using a short section of DNA from a specfic gene or genes. It is fast and accurate method that make ecological system more accessible by using short section or sequence of DNA instead of whole genome.

Dna Barcoding

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  It help in determine the species of a cell line. It involve amplifying and sequencing mitochondrial DNA because mitochondrial DNA shows 10-15 folds more difference between species than nuclear DNA. The Genetic analysis of mitochondrial DNA is simpler than nuclear DNA because only single PCR and sequencing reaction is required mitochondrial DNA shows only very low within-species variation. The short sequence is generated from predefined region in genome called as markers. It is specfic for species for eg. CO1 or cox1 cytochrome c oxidase, 16s in animals , matK for plants ,Internal Trancribed Spacer(ITS) for the fungus and 18s for protists.
• It first time came in attention to researchers community in 2003 when a group of researchers published a paper titled “Biological identification through DNA barcodes” under Paul Hebert at university of Guelph. It is not a new concept as Carl Woese used molecular markers like rDNA and mtDNA and rRNA to identified archea i.e. prokaryotes but its advantage is it use short DNA section instead of whole genome and use for eukaryotes. That short DNA sequence or section taken from standard region of genome to generate DNA barcode. DNA barcode is made of four nucleotide base A(Adenine), G(Guanine),C(Cytosine) and T(Thymine). Each base represent by different colour in barcode as shown in fig.below .Each individual has uniquely identify sequence and vary from species to species .

SIGNIFICANCE OF DNA BARCODING IN ANIMAL CELL LINE CULTURE

• In animal cell line culture it is essential to have knowledge of genome of particular cell line used. Rather then study of whole genome, barcoding technique make it easier by only study of short section in identification and authentication of cell line. From this method unknown cell line can also be identified by comparing with existing barcodes. For animals, the most widely used barcodes is mitochondrial cytochrome C oxidase I (COI) locus, Cytb, 12S,or 16S.
• For reference various Libraies are formed this liberaries used for identification of cell line, also called as annotation, of sequences obtain from barcoding or metabarcoding. This data base assigned to previously identified cell lines. Eg BOLD, UNITE, Diat.barcode etc.
• Thus by identify the cell line by this method we can get information about the cell line can do do require modification, desired product , perform certain test on them. General procedure is explaining below.

METHODOLOGY of DNA BARCODING

1. Tiny fragment of sample used to extract the mitochondrial DNA or direct already extracted sample can also be used.

2. The obtain DNA sample undergoes PCR amplification and sequencing, and the resultant data is compared to a DNA sequence database to identify the cell line.

3. Tiny fragment of sample used to extract the mitochondrial DNA or direct already extracted sample can also be used.

4. The obtain DNA sample undergoes PCR amplification and sequencing, and the resultant data is compared to a DNA sequence database to identify the cell line

5. Degenerate primers are used for amplifying the barcodes region of mitochondrial Cytochrome C Oxidase1,and public sequence databases can be used to assign an identity.

6. The barcodes sequence may be placed in BOLD(Barcode Of Life Data System) for the convenience of future researchers , a reference library of DNA barcodes thus created.

METHODOLOGY OF DNA BARCODING

SIGNIFICANCE of DNA BARCODING

• Authentication of cell line

• Help in identification of species

• Prevent from cross contamination during cell culture

• Protect data of endangered species

• Identification of disease vector

• Identification of all life stages

ADAVANTAGES OF DNA BARCODING

• It is faster than traditional method of identification and allows the processing of multiple samples

• It provide us taxanomic resolution.

• It help in study of population genetics.

• It help in conservation of various ecological benfical species.\

• Help in controlling agricultural pests.

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