Multiple displacement amplification prior to single nucleotide polymorphism genotyping in epidemiologic studies.
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.
Tranah
Gregory J
GJ
Department of Epidemiology, Harvard School of Public Health, Boston, MA 02115, USA.
Lescault
Pamela J
PJ
Hunter
David J
DJ
De Vivo
Immaculata
I
D001483:Base Sequence; D016022:Case-Control Studies; D004247:DNA / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000235:genetics; D015340:Epidemiologic Research Design; D020869:Gene Expression Profiling / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D009061:Mouth Mucosa / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
D005819:Genetic Markers; D004247:DNA