Abstract
Spots of blood are routinely collected from newborn babies onto filter paper called Guthrie cards and used to screen for metabolic and genetic disorders. The archived dried blood spots are an important and precious resource for genomic research. Whole genome amplification of dried blood spot DNA has been used to provide DNA for genome-wide SNP genotyping. Here we describe a 96 well format procedure to extract DNA from a portion of a dried blood spot that provides sufficient unamplified genomic DNA for genome-wide single nucleotide polymorphism (SNP) genotyping. We show that SNP genotyping of the unamplified DNA is more robust than genotyping amplified dried blood spot DNA, is comparable in cost, and can be done with thousands of samples. This procedure can be used for genome-wide association studies and other large-scale genomic analyses that require robust, high-accuracy genotyping of dried blood spot DNA.
Highlights
Newborn screening programs around the world use Guthrie filter cards to preserve several spots of blood obtained from heel pricks of newborn infants
The original protocol for newborn screening of phenylketonuria utilized a portion of a dried blood spots (DBS) for the test [1], and DBS are currently used to screen for many diseases, technical advances permit all of these tests to be conducted on a similar portion of a DBS
Many PCRbased amplification analyses have shown that DNA fragments of hundreds to thousands of base pairs can be amplified from genomic DNA (gDNA) extracted from a small portion of a DBS [3,4,5] and probed for single nucleotide polymorphisms (SNPs) or gene deletions [6,7]
Summary
Newborn screening programs around the world use Guthrie filter cards to preserve several spots of blood obtained from heel pricks of newborn infants. These samples are used to screen infants for metabolic and genetic diseases such as phenylketonuria and cystic fibrosis, and they are stored as dried blood spots (DBS). It was shown that targeted SNP analysis can be done on whole genome amplified DNA (wgaDNA) from DBS [8,9,10,11,12] and that wgaDNA from DBS can be used for arraybased genome-wide SNP genotyping [13,14,15,16]
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