Construction of larger-size sequencing templates from degraded DNA

We describe a modified recombinant PCR procedure that allows the constructi on of longer fragments of DNA from shorter overlapping PCR products. This m ethod permits the construction of optimal-size templates for DNA sequence a nalysis (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 Jive overlappi ng amplification products (159, 126, 131, 131 and 93 bp) derived from a Pal eolithic tooth specimen.