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A powerful and rapid approach to human genome scanning using small quantities of genomic DNA

  • MARIAN BEEKMAN (a1) (a2), NICO LAKENBERG (a1), STACEY S. CHERNY (a3), PETER DE KNIJFF (a2), C. CORNELIS KLUFT (a1), GERT-JAN B. VAN OMMEN (a2), GEORGE P. VOGLER (a4), RUNE R. FRANTS (a2), DORRET I. BOOMSMA (a5) and P. ELINE SLAGBOOM (a1) (a6)...

Abstract

Dense maps of short-tandem-repeat polymorphisms (STRPs) have allowed genome-wide searches for genes involved in a great variety of diseases with genetic influences, including common complex diseases. Generally for this purpose, marker sets with a 10 cM spacing are genotyped in hundreds of individuals. We have performed power simulations to estimate the maximum possible inter-marker distance that still allows for sufficient power. In this paper we further report on modifications of previously published protocols, resulting in a powerful screening set containing 229 STRPs with an average spacing of 18·3 cM. A complete genome scan using our protocol requires only 80 multiplex PCR reactions which are all carried out using one set of conditions and which do not contain overlapping marker allele sizes. The multiplex PCR reactions are grouped by sets of chromosomes, which enables on-line statistical analysis of a set of chromosomes, as sets of chromosomes are being genotyped. A genome scan following this modified protocol can be performed using a maximum amount of 2·5 μg of genomic DNA per individual, isolated from either blood or from mouth swabs.

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Copyright

Corresponding author

Corresponding author. e-mail: P.Slagboom@PG.TNO.NL

A powerful and rapid approach to human genome scanning using small quantities of genomic DNA

  • MARIAN BEEKMAN (a1) (a2), NICO LAKENBERG (a1), STACEY S. CHERNY (a3), PETER DE KNIJFF (a2), C. CORNELIS KLUFT (a1), GERT-JAN B. VAN OMMEN (a2), GEORGE P. VOGLER (a4), RUNE R. FRANTS (a2), DORRET I. BOOMSMA (a5) and P. ELINE SLAGBOOM (a1) (a6)...

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