Construction of larger-size sequencing templates from degraded DNA.
We describe a modified recombinant PCR procedure that allows the construction of longer fragments of DNA from shorter, overlapping PCR products. This method permits the construction of optimal-size templates for DNA sequence analysis (500-600 bp) from DNA that is too degraded for direct amplification of fragments of this size. As an example, the entire hypervariable region 1 of human mtDNA (418 bp) was reconstructed in one step from five overlapping amplification products (159, 126, 131, 131 and 93 bp) derived from a Paleolithic tooth specimen.
Nasidze
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I
Max Planck Institute for Evolutionary Anthropology, Department of Evolutionary Genetics, Leipzig, Germany. nasidze@eva.mpg.de
Stoneking
M
M
D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000737:chemistry; D004274:DNA, Recombinant; D006801:Humans; D008969:Molecular Sequence Data; D010161:Paleodontology; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013698:Templates, Genetic; D014070:Tooth / Q000737:chemistry
D004272:DNA, Mitochondrial; D004274:DNA, Recombinant