12. Arrayed Primer Extension Reaction for Genotyping on Oligonucleotide Microarray
| Abstract |
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Arrayed primer extension reaction (APEX) is a straightforward and robust enzymatic genotyping method in which hundreds to
thousands of variations in the genome are simultaneously analyzed in a single multiplexed reaction. It differs from allele-specific
hybridization in that the genotype information in APEX is obtained by single base extension, performed by a specific DNA polymerase,
together with four different dye terminators. This approach ensures highly specific discrimination without allele-specific
hybridization, because the primer to be extended anneals just adjacent to the DNA base that needs to be identified. Selection
of primers for specific sites or their consecutive placement in tiled format, shifting them by one base, permits single-nucleotide
polymorphism analysis, mutation detection, or resequencing of the DNA template. Careful primer design also permits the analysis
of insertions, deletions, splice variants, gene copy numbers, or CpG islands within the genome for gene methylation studies,
by performing additional bisulfite reactions.
Affiliation(s): (4) Department of Biotechnology, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia
(5) The Estonian Biocentre, Tartu, Estonia
(6) MDC of United Laboratories of the Tartu University Hospital, Tartu, Estonia
(7) Estonian Genome Project Foundation, Tartu, Estonia
(5) The Estonian Biocentre, Tartu, Estonia
(6) MDC of United Laboratories of the Tartu University Hospital, Tartu, Estonia
(7) Estonian Genome Project Foundation, Tartu, Estonia
Book Title: Prenatal Diagnosis
Series: Methods in Molecular Biology | Volume: 444 | Pub. Date: May-09-2008 | Page Range: 161-167 | DOI: 10.1007/978-1-59745-066-9_12
Subject: Molecular Medicine
Key Words: Arrayed primer extension reaction (APEX) - genotyping - minisequencing - molecular diagnostics - mutation analysis - oligonucleotide microarray - resequencing
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