Primers for a PCR-Based Approach to Mitochondrial Genome Sequencing in Birds and Other Vertebrates

• Published in: Molecular phylogenetics and evolution Vol. 12; no. 2; pp. 105 - 114
• Main Authors: Sorenson, Michael D., Ast, Jennifer C., Dimcheff, Derek E., Yuri, Tamaki, Mindell, David P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.1999
• Subjects: Animals; Base Sequence; Biologi med inriktning mot molekylär evolution; Biologi med inriktning mot systematik; Biology with specialization in Molecular Evolution; Biology with specialization in Systematics; Birds - classification; Birds - genetics; Conserved Sequence - genetics; DNA - genetics; DNA Primers - genetics; DNA, Mitochondrial - genetics; Genetic Variation; Genome; Polymerase Chain Reaction - methods; Pseudogenes - genetics; Sequence Analysis, DNA - methods; Temperature; Templates, Genetic
• ISSN: 1055-7903; 1095-9513; 1095-9513
• DOI: 10.1006/mpev.1998.0602

A PCR-based approach to sequencing complete mitochondrial genomes is described along with a set of 86 primers designed primarily for avian mitochondrial DNA (mtDNA). This PCR-based approach allows an accurate determination of complete mtDNA sequences that is faster than sequencing cloned mtDNA. The primers are spaced at about 500-base intervals along both DNA strands. Many of the primers incorporate degenerate positions to accommodate variation in mtDNA sequence among avian taxa and to reduce the potential for preferential amplification of nuclear pseudogenes. Comparison with published vertebrate mtDNA sequences suggests that many of the primers will have broad taxonomic utility. In addition, these primers should make available a wider variety of mitochondrial genes for studies based on smaller data sets.