One-way sequencing of multiple amplicons from tandem repetitive mitochondrial DNA control region

Abstract

Repetitive DNA sequences not only exist abundantly in eukaryotic nuclear genomes, but also occur as tandem repeats in many animal mitochondrial DNA (mtDNA) control regions. Due to concerted evolution, these repetitive sequences are highly similar or even identical within a genome. When long repetitive regions are the targets of amplification for the purpose of sequencing, multiple amplicons may result if one primer has to be located inside the repeats. Here, we show that, without separating these amplicons by gel purification or cloning, directly sequencing the mitochondrial repeats with the primer outside repetitive region is feasible and efficient. We exemplify it by sequencing the mtDNA control region of the mosquito Aedes albopictus, which harbors typical large tandem DNA repeats. This one-way sequencing strategy is optimal for population surveys.

Keywords::

• Aedes albopictus
• concerted evolution
• repetitive DNA
• sequencing strategy

Acknowledgements

This work was funded by Cooperative Agreement between USDA-ARS and Rutgers University (USDA-ARS-58-6615-8-105), which is entitled “Area-wide Management of the Asian Tiger Mosquito.”

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.