The first description of a methodology for sequencing the entire mtGenome was by Deborah Nickerson's group at the University of Washington (Rieder et al. 1998). They used 24 pairs of primers to amplify PCR products ranging in size from 765 bp to 1162 bp. These primer pairs provide on average almost 200 bases of overlap between the various PCR products spanning the mtGenome. Ingman et al. (2000) used the Nickerson laboratory sequencing strategy to launch the era of mitochondrial population genomics when they sequenced 53 mtGenomes from diverse world population groups. Max Ingman maintains an mtGenome polymorphism database at http://www.genpat.uu.se/mtDB/.
In the past few years, a number of other methodologies have appeared in the literature for sequencing entire mtGenomes (Table 10.7). Regardless of the sequencing strategy used, the biggest challenge in conducting this work remains efforts to reduce and eliminate errors in sequence review (see Herrnstadt et al. 2003). Fortunately, the reference sequence (rCRS) was updated prior to the explosion of mtGenome information that began with Ingman et al. (2000).
The program MitoAnalyzer (Lee and Levin 2002) can be used to evaluate the location of an observed polymorphic nucleotide in the mtGenome. MitoAnalyzer is available online at http://www.cstl.nist.gov/biotech/strbase/ mitoanalyzer.html.
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