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Journal of Lipid Research, Vol 33, 1563-1567, Copyright © 1992 by Lipid Research, Inc.

ARTICLES

Apolipoprotein B(Arg3500----Gln) allele specific polymerase chain reaction: large-scale screening of pooled blood samples

V Ruzicka, W Marz, A Russ and W Gross
Gustav Embden-Centre of Biological Chemistry, Johann Wolfgang Goethe- University, University Hospital, Frankfurt am Main, Germany.

A two-step polymerase chain reaction (PCR) method for the rapid detection of the apolipoprotein B(Arg3500----Gln) mutation in a mixture of pooled blood samples is described. In the first step PCR, a short gene fragment surrounding codon 3500 is amplified. Subsequently the reaction product is subjected to a second amplification in which a mutation-specific primer is used. A PCR product is generated only if the mutant sequence is present in the DNA pool. Individuals carrying the mutation can then be identified by PCR with mutagenic primers and MspI restriction typing, essentially as described by Hansen et al. (J. Lipid Res. 1991. 32: 1229-1233).
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