A low-cost, high-throughput, automated single nucleotide polymorphism assay for forensic human DNA applications.
Single nucleotide polymorphism (SNP) analysis of human DNA for the purpose of identification has some promising attributes. The question of approach is critical to the eventual adoption of this technology. The use of a low-volume open array platform was tested with a small selected set of eight SNP primers that have a low F(ST) (the proportion of the total genetic variance contained in a subpopulation [S subscript] relative to the total genetic variance [T subscript]) in human populations. Because multiple SNPs must be interrogated, issues concerning DNA concentration, total DNA, and whole genome amplification were investigated. Excellent correlations were obtained for seven of the eight SNP assays on a set of DNA samples of known configuration over a broad concentration range spanning 25-150ng/microl in blind studies. These seven SNP assays were then applied to 39 DNA samples in a population from southern India. These SNPs were sufficient to individualize each member of this sample population. In a paternity study, these same SNPs showed clear parental relationships. For low amounts of genomic DNA, the use of a commercially available whole genome amplification kit showed promise for genotyping sub-nanogram samples. Discrimination against nonhuman DNA was also demonstrated successfully. Because of the very low quantities of reagents used in the assay, the cost per test becomes reasonably inexpensive. Overall, using commercially available SNP assays, the OpenArray platform showed excellent promise as a highly automated, low-volume, high-throughput system for SNP analysis with potential applications to relevant forensic analyses such as identification and paternity.
Pomeroy
Robert
R
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, 92093, USA. rpomeroy@ucsd.edu
Duncan
George
G
Sunar-Reeder
Bulbin
B
Ortenberg
Elen
E
Ketchum
Melba
M
Wasiluk
Hannah
H
Reeder
Dennis
D
D000818:Animals; D002415:Cats; D003365:Costs and Cost Analysis; D004247:DNA / Q000097:blood; D053836:DNA Degradation, Necrotic; D018732:Forensic Anthropology; D053803:Forensic Genetics / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D014644:Genetic Variation; D015894:Genome, Human; D005838:Genotype; D006197:Hair; D006801:Humans; D008832:Microchemistry; D021141:Nucleic Acid Amplification Techniques; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D010334:Paternity; D020641:Polymorphism, Single Nucleotide; D012984:Software; D013995:Time
D004247:DNA