ADLAPH: A molecular haplotyping method based on allele-discriminating long-range PCR.
We present a method, called Allele-Discriminating Long and Accurate PCR Haplotyping (ADLAPH), for directly determining haplotypes from an extended genomic region. This method uses allele-discriminating primers in long-range PCR to amplify only one of the two chromosome homologues for the region of interest. Haplotypes are then determined from these phase-separated PCR fragments by conventional single nucleotide polymorphism (SNP) genotyping methods. This simple robust procedure makes it practical for high-throughput haplotyping of unrelated individuals, and potentially allows direct observation of haplotype information for up to 40 kb or more. We demonstrate the feasibility of this molecular haplotyping procedure by generating apolipoprotein E (APOE) haplotypes from 100 unrelated subjects.
Yu
Chang-En
CE
Geriatric Research, Education, and Clinical Center, Veterans Affairs Puget Sound Health Care System, Seattle, WA 98108, USA. changeyu@u.washington.edu
Devlin
Bernie
B
Galloway
Nichole
N
Loomis
Elaine
E
Schellenberg
Gerard D
GD
D000483:Alleles; D001057:Apolipoproteins E / Q000235:genetics; D001483:Base Sequence; D001499:Bayes Theorem; D002875:Chromosomes / Q000235:genetics*; D017931:DNA Primers; D005838:Genotype; D006239:Haplotypes / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA
D001057:Apolipoproteins E; D017931:DNA Primers