title: Evaluating whole genome amplification via multiply-primed rolling circle amplification for SNP genotyping of samples with low DNA yield.
abstract: The amount of available DNA is often a limiting factor in pursuing genetic analyses of large-scale population cohorts. An association between higher DNA yield from blood and several phenotypes associated with inflammatory states has recently been demonstrated, suggesting that exclusion of samples with very low DNA yield may lead to biased results in statistical analyses. Whole genome amplification (WGA) could present a solution to the DNA concentration-dependent sample selection. The aim was to thoroughly assess WGA for samples with low DNA yield, using the multiply-primed rolling circle amplification method. Fifty-nine samples were selected with the lowest DNA yield (less than 7.5 microg) among 799 samples obtained for one population cohort. The genotypes obtained from two replicate WGA samples and the original genomic DNA were compared by typing 24 single nucleotide polymorphisms (SNPs). Multiple genotype discrepancies were identified for 13 of the 59 samples. The largest portion of discrepancies was due to allele dropout in heterozygous genotypes in WGA samples. Pooling the WGA DNA replicates prior to genotyping markedly improved genotyping reproducibility for the samples, with only 7 discrepancies identified in 4 samples. The nature of discrepancies was mostly homozygote genotypes in the genomic DNA and heterozygote genotypes in the WGA sample, suggesting possible allele dropout in the genomic DNA sample due to very low amounts of DNA template. Thus, WGA is applicable for low DNA yield samples, especially if using pooled WGA samples. A higher rate of genotyping errors requires that increased attention be paid to genotyping quality control, and caution when interpreting results.
authors.affiliation: Department of Molecular Medicine, National Public Health Institute, Helsinki, Finland. kaisa.silander@ktl.fi
authors.forename: Kaisa
authors.initials: K
authors.lastname: Silander
authors.forename: Kati
authors.initials: K
authors.lastname: Komulainen
authors.forename: Pekka
authors.initials: P
authors.lastname: Ellonen
authors.forename: Minttu
authors.initials: M
authors.lastname: Jussila
authors.forename: Mervi
authors.initials: M
authors.lastname: Alanne
authors.forename: Minna
authors.initials: M
authors.lastname: Levander
authors.forename: Päivi
authors.initials: P
authors.lastname: Tainola
authors.forename: Kari
authors.initials: K
authors.lastname: Kuulasmaa
authors.forename: Veikko
authors.initials: V
authors.lastname: Salomaa
authors.forename: Markus
authors.initials: M
authors.lastname: Perola
authors.forename: Leena
authors.initials: L
authors.lastname: Peltonen
authors.forename: Janna
authors.initials: J
authors.lastname: Saarela
mesh_terms: D004247:DNA / Q000032:analysis*; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D020641:Polymorphism, Single Nucleotide; D011786:Quality Control; D015203:Reproducibility of Results
chemical_list: D004247:DNA