A whole genome amplification method to generate long fragments from low quantities of genomic DNA

Ralf Kittler, Mark Stoneking, Manfred Kayser

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)237-244
Number of pages8
JournalAnalytical biochemistry
Volume300
Issue number2
DOIs
StatePublished - Jan 15 2002

Keywords

  • DOP-PCR
  • Degenerate oligonucleotide-primed PCR
  • Microsatellites
  • STR
  • Short tandem repeats
  • Whole genome amplification

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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