| Title : Molecular haplotyping by linking emulsion PCR: analysis of paraoxonase 1 haplotypes and phenotypes - Wetmur_2005_Nucleic.Acids.Res_33_2615 |
| Author(s) : Wetmur JG , Kumar M , Zhang L , Palomeque C , Wallenstein S , Chen J |
| Ref : Nucleic Acids Research , 33 :2615 , 2005 |
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Abstract :
Linking emulsion PCR (LE-PCR) enables formation of minichromosomes preserving phase information of two polymorphic loci, hence the haplotype. Emulsion PCR confines two amplicons of two linked polymorphic sites on a single template molecule to one aqueous-phase droplet. Linking PCR uses biotinylated, overlapping linking primers to connect these amplicons in the droplet. After LE-PCR, unlinked amplicons are removed on streptavidin-coated magnetic beads and single-stranded runoff products are capped by primer extension. Quantitative ASPCR can then be used to ascertain the haplotypes of the two polymorphic loci on the minichromosomes. Using LE-PCR, we determined the human paraoxonase-1 [PON1] molecular haplotypes at three loci (-909g>c, L55M, Q192R) in women who were compound heterozygotes for -909g>c/L55M (n = 89), -909g>c/Q192R (n = 77) and L55M/Q192R (n = 68). We observed a strong association between PON1 substrate specificity (paraoxon/phenylacetate substrate activity ratios) and -909g>c/Q192R haplotype. We have demonstrated here a powerful molecular haplotyping technology that can be applied in population studies. |
| PubMedSearch : Wetmur_2005_Nucleic.Acids.Res_33_2615 |
| PubMedID: 15886392 |
Wetmur JG, Kumar M, Zhang L, Palomeque C, Wallenstein S, Chen J (2005)
Molecular haplotyping by linking emulsion PCR: analysis of paraoxonase 1 haplotypes and phenotypes
Nucleic Acids Research
33 :2615
Wetmur JG, Kumar M, Zhang L, Palomeque C, Wallenstein S, Chen J (2005)
Nucleic Acids Research
33 :2615