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Ancient DNA (a-DNA) studies in past three decades has been uplifted from short stretch archaeological mtDNA retrieval to reconstruction of complete mitochondrial genome sequence using next generation sequencing. Study of a-DNA acts as validating tool to test the assumed hypothesis used in origin, evolution, diversification, distribution, domestication and extinction of species; although, scientific progression in the field is frequently hindered by erroneous reports due to contamination and other technical bottlenecks. With the advent of appropriate methodologies such as 3’-C3 spacer tagged oligonucleotide blocking, shotgun sequencing, amplicon sequencing Polymerase Chain Reaction, single primer extension methodology, development of broad spectrum ancient sequence primer, slippage proof amplification methods, use of multiplexing strategy in PCR (Polymerase Chain Reaction) amplification, etc. in a-DNA research try to minimize the difficulties associated with it. Present review highlighted the pitfalls as well as prospects of a-DNA study and tries to assess the significance and acceptability of a-DNA research for its future application in understanding of evolutionary biology.

Keywords

a-DNA, Extraction Difficulties, Modern Technologies, Retrospect of a-DNA
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