INVERSE POLYMERASE CHAIN REACTION Meaning and
Definition
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Inverse Polymerase Chain Reaction (Inverse PCR) is a molecular biology technique used to amplify DNA sequences adjacent to a known DNA fragment or gene of interest by reversing the traditional implementation of the Polymerase Chain Reaction (PCR) method. It is commonly employed to identify unknown sequences flanking a known DNA piece, and to facilitate the analysis of gene rearrangements, deletions, insertions, or translocations within the genome.
The process of Inverse PCR involves several steps. Initially, the target DNA fragment is circularized by ligation, which results in a closed loop. Subsequently, PCR amplification is performed using primers facing outward from the known DNA sequence or gene of interest. The amplification process proceeds from the known region toward the unknown region, amplifying the DNA located adjacent to the known sequence. These newly amplified fragments are then available for further analysis, which comprises sequencing, mapping, or cloning.
Inverse PCR provides an opportunity to investigate aspects of the genome which were previously inaccessible due to their unknown compositions. It is particularly useful when working with limited or unknown genomic DNA sequences, as it allows for the exploration of sequences adjacent to known regions. This technique is frequently employed in molecular biology research, contributing to a better understanding of gene structure, gene regulation, and the identification of important DNA elements in biological systems.
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