A whole genome amplification method to generate long fragments from low quantities of genomic DNA.
Several whole genome amplification strategies have been developed to preamplify the entire genome from minimal amounts of DNA for subsequent molecular genetic analysis. However, none of these techniques has proven to amplify long products from very low (nanogram or picogram) quantities of genomic DNA. Here we report a new whole genome amplification protocol using a degenerate primer (DOP-PCR) that generates products up to about 10 kb in length from less than 1 ng genomic template DNA. This new protocol (LL-DOP-PCR) allows in the subsequent PCR the specific amplification, with high fidelity, of DNA fragments that are more than 1 kb in length. LL-DOP-PCR provides significantly better coverage for microsatellites and unique sequences in comparison to a conventional DOP-PCR method.
Kittler
Ralf
R
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Inselstrasse 22, D-04103 Leipzig, Germany.
Stoneking
Mark
M
Kayser
Manfred
M
D004247:DNA / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D015894:Genome, Human; D006801:Humans; D008297:Male; D008832:Microchemistry / Q000379:methods; D018895:Microsatellite Repeats / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D013698:Templates, Genetic; D013997:Time Factors
D017931:DNA Primers; D004247:DNA