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PMID: 12889810 Published · ppublish English Evaluation Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Validation Study

Multiple displacement amplification prior to single nucleotide polymorphism genotyping in epidemiologic studies.

Biotechnology letters ·Vol. 25 ·No. 13 ·2003-07-00 ·Pages 1031-6

Tranah GJ, Lescault PJ, Hunter DJ, De Vivo I

Abstract

We assessed the whole genome amplification strategy, known as multiple displacement amplification (MDA), for use with the TaqMan genotyping platform for DNA samples derived from two case-control studies nested in the Nurses' Health Study and the Physicians' Health Study. Our objectives were to (1) quantify DNA yield from samples of varying starting concentrations and (2) assess whether MDA products give an accurate representation of the original genomic sequence. Multiple displacement amplification yielded a mean 23000-fold increase in DNA quantity and genotyping results demonstrate 99.95% accuracy across six SNPs from four genes for 352 samples included in this study. These results suggest that MDA will provide a sufficiently robust amplification of limiting samples of genomic DNA that can be used for SNP genotyping in large case-control studies of complex diseases.

MeSH Terms
Base Sequence Case-Control Studies DNA/analysis,blood,chemistry,genetics Epidemiologic Research Design Gene Expression Profiling/methods Genetic Markers/genetics Genome, Human Genotype Humans Molecular Sequence Data Mouth Mucosa/chemistry Nucleic Acid Amplification Techniques/methods Polymorphism, Single Nucleotide/genetics Reproducibility of Results Sensitivity and Specificity Sequence Analysis, DNA/methods
Chemicals
Genetic Markers DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tranah Gregory J
Department of Epidemiology, Harvard School of Public Health, Boston, MA 02115, USA.
Lescault Pamela J
Hunter David J
De Vivo Immaculata
Article Info
Journal
Biotechnology letters
Abbr.
Biotechnol Lett
ISSN
0141-5492
Published
2003-07-00
Pages
1031-6
Language
English
Region
Netherlands
NLM ID
8008051
Subset
IM
Grants
NCI NIH HHS · CA49449 · United States
NCI NIH HHS · CA65725 · United States
NCI NIH HHS · CA82838 · United States
NCI NIH HHS · CA87969 · United States
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